Haplotypes Associated with Improved Dicamba Tolerance and Glyphosate Tolerance in Transgenic Soybean Plants

ABSTRACT

The present invention provides methods and compositions for the identification and selection of loci modulating phenotypic expression of an herbicide tolerance trait in plant breeding. In addition, methods are provided for screening germplasm entries for the performance and expression of this trait.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No. 14/402,700, filed Nov. 21, 2014, now U.S. Pat. No. 10,604,767, which is a national stage application, filed under 35 U.S.C. § 371, of International Patent Application No. PCT/US2013/042349, filed May 23, 2013, which claims the benefit of U.S. Provisional Patent Application No. 61/779,739, filed Mar. 13, 2013; U.S. Provisional Patent Application No. 61/753,725, filed Jan. 17, 2013; U.S. Provisional Patent Application No. 61/753,693, filed Jan. 17, 2013; U.S. Provisional Patent Application No. 61/650,869, filed May 23, 2012; and U.S. Provisional Patent Application No. 61/650,852, filed May 23, 2012, each of which are incorporated by reference herein in its entirety.

INCORPORATION OF SEQUENCE LISTING

A sequence listing containing the file named “46_21_59246 A_PCT.txt” which is 33,292 bytes (measured in MS-Windows®) and created on May 20, 2013, comprises 56 nucleotide sequences, is provided herewith via the USPTO's EFS system and is herein incorporated by reference in its entirety.

INCORPORATION OF TABLE 2

A listing of various soybean linkage group L (chromosome 19) markers is provided herewith in the Specification as Table 2.

BACKGROUND

International Patent Application Publication WO 2012/031097 describes genetic regions of soybean linkage group L that contain polymorphic loci that are associated with an undesirable “yellow flash” phenotype that is observed in the foliage of certain soybean varieties that comprise a transgene that confers resistance to glyphosate that are exposed to glyphosate.

SUMMARY

“Dicamba intolerance” is an undesirable phenotype observed in certain soybean varieties that comprise a transgene that can confer resistance to the broad-spectrum herbicide dicamba. After application of dicamba, it has been discovered that the leaves of certain soybean plant varieties comprising the transgene that confers resistance to dicamba can exhibit a “dicamba intolerance phenotype” comprising malformation (epinasty) of the main stem and petioles upon exposure to dicamba. The epinastic growth habit of such “dicamba intolerant” transgenic plants is manifest in pronounced bending/twisting of the main stem and petioles. In dicamba intolerant transgenic soybean plants exposed to dicamba, the upper nodes and petioles may die, but lower portion of the plant may remain vegetative and new growth can be limited. However, other soybean plant varieties containing the same transgene that confers resistance to dicamba do not exhibit the dicamba intolerance phenotype when co-exposed to the same dosage of dicamba. The dicamba intolerance phenotype can be observed within approximately 2 to 10 days after herbicide application in certain soybean varieties comprising the transgene that confers resistance to dicamba. The dicamba intolerance phenotype is undesirable as it can lead to reduced yield in certain transgenic soybean plant varieties exposed to dicamba.

Although the dicamba intolerance phenotype can be observed within approximately 2 to 10 days after dicamba application in certain soybean varieties comprising the transgene that confers dicamba resistance, distinct soybean varieties that comprise the same dicamba resistance transgene integrated at the same chromosomal locus (i.e. the same transgenic event) can show various degrees of dicamba intolerance upon exposure to high doses of dicamba. Some varieties comprising the dicamba resistance transgene insertion are highly tolerant to high dosages of dicamba, showing no dicamba intolerance phenotype (i.e. a “dicamba tolerance phenotype”), while other varieties comprising the same dicamba resistance transgene insertion are highly susceptible to high dosages of dicamba, showing a severe dicamba intolerance phenotype. Provided herein are soybean plants comprising an introgressed genomic region associated with a dicamba tolerance phenotype. Also provided herein are markers that reside outside of a genomic region associated with a dicamba tolerance phenotype and that facilitate breeding activities that include, but are not limited to, introgression of this genomic region. Markers and specific alleles thereof that are associated with a dicamba tolerance phenotype are also provided. Methods of obtaining a soybean plant that exhibits a dicamba tolerance phenotype and methods of obtaining a soybean plant comprising in its genome at least one dicamba tolerance locus are also provided. Methods that provide for the introgression of a genomic region associated with a dicamba tolerance phenotype into soybean germplasm that has a genomic region associated with a dicamba intolerance phenotype are also provided. Identification of molecular markers associated with loci that confer the dicamba tolerance phenotype has significant economic value. By using markers associated with the dicamba tolerance trait, breeders can select soybean varieties with the favorable alleles (i.e. alleles that are not associated with the dicamba intolerance trait) for use in trait integration. They can also use the markers to help them eliminate unfavorable alleles (i.e. alleles that are associated with the dicamba intolerance trait) in soybeans. In certain embodiments, commercially desirable transgenic soybean lines that carry a genomic region that is associated with a “dicamba tolerance” phenotype and tolerate high dosages of dicamba are thus provided.

It has also been surprisingly observed that soybean plants comprising the dicamba tolerance loci, a transgene conferring resistance to dicamba, and a transgene conferring resistance to glyphosate also exhibit improved reproductive tolerance to glyphosate application relative to plants with the same two transgenes that lack the dicamba tolerance loci. Although the glyphosate reproductive intolerance phenotype can be observed after late stage (i.e. V6/R1) glyphosate application in certain soybean varieties comprising the transgenes that confer dicamba and glyphosate resistance, distinct soybean varieties that comprise the same dicamba and glyphosate resistance transgene integrated at the same chromosomal loci (i.e. the same transgenic events) can show various degrees of glyphosate reproductive intolerance (i.e. varying degrees of sterility) upon such exposure to glyphosate. Some varieties comprising the dicamba and glyphosate resistance transgene insertions are highly tolerant to late stage glyphosate application, showing no sterility phenotype (i.e. a “glyphosate reproductive intolerance phenotype”), while other varieties comprising the same dicamba and glyphosate resistance transgene insertions are highly susceptible to late stage glyphosate application, showing varying levels of sterility. Provided herein are soybean plants comprising an introgressed genomic region associated with a dicamba tolerance phenotype that also provide for reproductive tolerance to glyphosate. Also provided herein are markers that reside outside of a genomic region associated with a dicamba tolerance/reproductive tolerance to glyphosate phenotype and that facilitate breeding activities that include, but are not limited to, introgression of this genomic region. Markers and specific alleles thereof that are associated with a dicamba tolerance/reproductive tolerance to glyphosate are also provided. Methods of obtaining a soybean plant that exhibits reproductive tolerance to glyphosate and methods of obtaining a soybean plant comprising in its genome at least one dicamba tolerance/reproductive tolerance to glyphosate locus are also provided. Methods that provide for the introgression of a genomic region associated with reproductive tolerance to glyphosate into soybean germplasm that has a genomic region associated with a reproductive tolerance to glyphosate are also provided. Identification of molecular markers associated with loci that confer reproductive tolerance to glyphosate has significant economic value. By using markers associated with the reproductive tolerance to glyphosate trait, breeders can select soybean varieties with the favorable alleles (i.e. alleles that are not associated with the glyphosate reproductive intolerance trait) for use in trait integration. They can also use the markers to help them eliminate unfavorable alleles (i.e. alleles that are associated with the glyphosate reproductive intolerance trait) in soybeans. In certain embodiments, commercially desirable transgenic soybean lines that carry a genomic region that is associated with a “glyphosate reproductive tolerance” phenotype and tolerate late stage (i.e. V6/R1) application of glyphosate are thus provided.

Methods of identifying a soybean plant that comprises a genotype associated with a dicamba tolerance phenotype and/or a glyphosate reproductive tolerance phenotype are thus provided.

In certain embodiments, the plurality of soybean plants comprises a population that is obtained by: i) crossing a parent plant comprising at least one dicamba tolerance locus with a parent plant comprising at least one dicamba intolerance locus; or, ii) obtaining seed or progeny from a parental plant segregating for at least one dicamba tolerance locus. In certain embodiments, the population contains plants that comprise a transgene that confers resistance to dicamba. In certain embodiments, the aforementioned methods can further comprise the step of assaying for the presence of at least one additional marker, where the additional marker is either linked or unlinked to the linkage group L genomic region. In certain embodiments of the aforementioned methods, the plurality of soybean plants, the soybean plant, and/or progeny thereof are exposed to a dosage of dicamba sufficient to cause dicamba intolerance in a susceptible variety. In certain embodiments of the aforementioned methods, a plant that exhibits a dicamba tolerance phenotype is selected.

Also provided herewith are methods for producing a soybean plant comprising in its genome at least one introgressed dicamba tolerance locus. Also provided herewith are soybean plants comprising an introgressed dicamba tolerance locus made by the aforementioned methods. In certain embodiments, a soybean plant comprising an introgressed dicamba tolerance locus and one or more polymorphic loci comprising alleles or combinations of alleles that are not found in a dicamba tolerant soybean variety and that are linked to the introgressed dicamba tolerance locus, where the plant is produced by the aforementioned methods are provided.

Also provided are soybean plants comprising an introgressed dicamba tolerance locus and one or more polymorphic loci comprising alleles or combinations of alleles that are not found in a dicamba tolerant soybean variety and that are linked to the introgressed dicamba tolerance locus.

Methods of identifying a soybean plant that comprises a genotype associated with dicamba tolerance and/or reproductive tolerance to glyphosate are thus provided. In certain embodiments, the methods can comprise detecting in a soybean plant an allele in at least one genetic locus associated with dicamba tolerance and/or reproductive tolerance to glyphosate, where the genetic locus is in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12), and denoting that the plant comprises a genotype associated with dicamba tolerance. In certain embodiments, the methods can further comprise the step of selecting the denoted plant from a population of plants. In certain embodiments, the plant comprises a transgene that confers resistance to dicamba and/or a transgene that confers resistance to glyphosate. In certain embodiments, the soybean plant or progeny thereof is exposed to a dosage of dicamba sufficient to cause a deleterious effect in a susceptible variety comprising the transgene and/or is exposed to a dosage of glyphosate sufficient to cause sterility in a susceptible variety comprising the transgene(s). In certain embodiments of any of the aforementioned methods, a plant that exhibits dicamba tolerance and/or reproductive tolerance to glyphosate is selected. In certain embodiments of any of the aforementioned methods, a genotype associated with a dicamba tolerance comprises at least one polymorphic allele of at least one marker in a first sub-region of the linkage group L region that is flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6) and/or at least one polymorphic allele of at least one marker in a second sub-region of the linkage group L region that is flanked by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12) and/or at least one polymorphic allele of at least one marker in a third sub-region of the linkage group L region that is flanked by loci BU55345 (SEQ ID NO:7) and M0114388 (SEQ ID NO:8) is provided. In certain embodiments of any of the aforementioned methods, the genotype associated with dicamba tolerance comprises at least one polymorphic allele of at least one marker in the linkage group L region selected from the group consisting of a TT allele M0205350 (SEQ ID NO: 10), a TT allele of M0101742 (SEQ ID NO: 5), a CC allele of M0102027 (SEQ ID NO: 11), and an AA allele of NGMAX008197032 (SEQ ID NO:52).

Methods for obtaining a soybean plant comprising in its genome at least one dicamba tolerance locus are also provided. In certain embodiments, these methods can compromise the steps of: (a) genotyping a plurality of soybean plants with respect to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12); and, (b) selecting a soybean plant comprising in its genome at least one genetic locus comprising a genotype associated with dicamba tolerance. In certain embodiments of these methods, the genotype associated with dicamba tolerance comprises at least one polymorphic allele of at least one marker in a first sub-region of the linkage group L region flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6) and/or at least one polymorphic allele of at least one marker in a second sub-region of the linkage group L region that is flanked by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12) and/or at least one polymorphic allele of at least one marker in a third sub-region of the linkage group L region that is flanked by loci BU55345 (SEQ ID NO:7) and M0114388 (SEQ ID NO:8). In certain embodiments of any of these aforementioned methods, the genotype associated with dicamba tolerance comprises at least one polymorphic allele of at least one marker in the first linkage group L region, the first sub-region, the second sub-region, or the third sub-region, where the marker is selected from the group consisting of a TT allele M0205350 (SEQ ID NO: 10), a TT allele of M0101742 (SEQ ID NO: 5), a CC allele of M0102027 (SEQ ID NO: 11), and an AA allele of NGMAX008197032 (SEQ ID NO:52). In certain embodiments of these methods, the plurality of soybean plants comprises a population that is obtained by: i) crossing a parent plant comprising at least one dicamba tolerance locus with a parent plant comprising at least one dicamba sensitivity locus; or, ii) obtaining seed or progeny from a parental plant segregating for at least one dicamba tolerance locus. In certain embodiments of these methods, the population contains plants that comprise at least one transgene that confers resistance to dicamba and/or a transgene that confers resistance to glyphosate. In certain embodiments of any of the aforementioned methods, the methods can further comprise the step of assaying for the presence of at least one additional marker, where the additional marker is either linked or unlinked to the linkage group L genomic region. In certain embodiments of any of the aforementioned methods, the plurality of soybean plants, the soybean plant, and/or progeny thereof are exposed to a dosage of dicamba sufficient to cause a deleterious effect in a susceptible variety comprising the transgene and/or is exposed to a dosage of glyphosate sufficient to cause sterility in a susceptible variety comprising the transgene. In certain embodiments of any of the aforementioned methods, a plant that exhibits dicamba tolerance and/or reproductive tolerance to glyphosate is selected.

Methods for producing a soybean plant comprising in its genome at least one introgressed dicamba tolerance locus are also provided. In certain embodiments, these methods comprise the steps of: (a) crossing a first soybean plant with a dicamba tolerance locus with a second soybean plant comprising: a dicamba sensitivity locus in a first linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6) and/or at least one polymorphic allele of at least one marker in a second sub-region of the linkage group L region that is flanked by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12) and/or at least one polymorphic allele of at least one marker in a third sub-region of the linkage group L region that is flanked by loci BU55345 (SEQ ID NO:7) and M0114388 (SEQ ID NO:8) and at least one linked polymorphic locus not present in the first soybean plant to obtain a population segregating for the dicamba tolerance loci and the linked polymorphic locus; (b) detecting at least two polymorphic nucleic acids in at least one soybean plant from the population, where at least one of the polymorphic nucleic acids is located in the first linkage group L region and/or the second linkage group L region and where at least one of the polymorphic amino acids is a linked polymorphic locus not present in the first soybean plant; and (c) selecting a soybean plant comprising a genotype associated with dicamba tolerance and at least one linked marker found in the second soybean plant comprising a dicamba sensitivity locus but not in the first soybean plant, thereby obtaining a soybean plant comprising in its genome at least one introgressed dicamba tolerance locus. In certain embodiments of these methods, at least one of the first or the second soybean plants comprises a transgene that confers resistance to dicamba and/or a transgene that confers resistance to glyphosate. In certain embodiments of these methods, the population, the selected soybean plant, and/or progeny of selected soybean plant is exposed to a dosage of dicamba sufficient to cause a deleterious effect in a susceptible variety comprising the transgene and/or is exposed to a dosage of glyphosate sufficient to cause sterility in a susceptible variety comprising the transgene. In certain embodiments of these methods, the polymorphic nucleic acid detected in step (b) is detected with at least one marker selected from the group consisting of M0205350 (SEQ ID NO: 10), M0101742 (SEQ ID NO: 5), M0102027 (SEQ ID NO: 11), and NGMAX008197032 (SEQ ID NO:52). In certain embodiments of these methods, the polymorphic nucleic acid detected in step (b) comprises a TT allele of M0205350 (SEQ ID NO: 10), a TT allele of M0101742 (SEQ ID NO: 5), a CC allele of M0102027 (SEQ ID NO: 11), and an AA allele of NGMAX008197032 (SEQ ID NO:52). In certain embodiments of these methods, the polymorphic nucleic acid detected in step (b) is detected with marker M0205350 (SEQ ID NO: 10) or M0102027 (SEQ ID NO: 11). In certain embodiments of these methods, the polymorphic nucleic acids are detected with marker M0101742 (SEQ ID NO: 5). In certain embodiments of these methods, the polymorphic nucleic acids are detected with marker NGMAX008197032 (SEQ ID NO:52). In certain embodiments of any of the aforementioned methods, the linked polymorphic locus is detected with a genotypic marker, a phenotypic marker, or both. In certain embodiments of these methods, the linked polymorphic locus is detected with a marker that is located within about 1000, 500, 100, 40, 20, 10, or 5 kilobases (Kb) of the dicamba tolerance locus. In certain embodiments of these methods, the linked polymorphic locus is detected with at least one marker selected from the group consisting of asmbl_11856 (SEQ ID NO: 1), TC122822 (SEQ ID NO: 2), BI967232 (SEQ ID NO: 3), M0205537 (SEQ ID NO: 15), M0202715 (SEQ ID NO: 16), M0206286 (SEQ ID NO: 17), M0206054 (SEQ ID NO: 18), and M0205375 (SEQ ID NO: 19).

Transgenic soybean plants comprising introgressed linkage group L regions comprising at least one polymorphic allele of at least one marker in a first sub-region of the linkage group L region that flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6) and/or at least one polymorphic allele of at least one marker in a second sub-region of the linkage group L region that is flanked by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12) and/or at least one polymorphic allele of at least one marker in a third sub-region of the linkage group L region that is flanked by loci BU55345 (SEQ ID NO:7) and M0114388 (SEQ ID NO:8), where the polymorphic alleles are associated with dicamba tolerance and/or reproductive tolerance to glyphosate, and where the plant comprises a transgene that confers resistance to dicamba are also provided. In certain embodiments, the polymorphic alleles comprise a TT allele of M0205350 (SEQ ID NO: 10), a TT allele of M0101742 (SEQ ID NO: 5), a CC allele of M0102027 (SEQ ID NO: 11), and an AA allele of NGMAX008197032 (SEQ ID NO:52). In certain embodiments, the transgenic plant exhibits dicamba tolerance. In certain embodiments, the transgenic plant further comprises a transgene that confers resistance to glyphosate and exhibits reproductive tolerance to glyphosate. In certain embodiments of any one of the aforementioned methods, the plant further comprises at least one of a 2,4-D, glufosinate, bromoxynil, acetolactate synthase (ALS), acetyl CoA carboxylase (ACCase), hydroxyphenyl pyruvate dioxygenase (HPPD), or sulfonylurea herbicide resistance transgenes and/or at least one transgene selected from the group of transgenes conferring insect resistance, nematode resistance, fungal resistance, an improvement in seed oil quantity, an improvement in seed oil quality, abiotic stress resistance, and intrinsic yield increases. In certain embodiments, the insect resistance conferring transgene is a transgene that expresses an insecticidal Bacillus thuringiensis protein.

Also provided herein are soybean plants comprising a dicamba tolerance locus, a transgene conferring resistance to glyphosate, a transgene conferring resistance to dicamba, where the plants exhibit both improved dicamba tolerance and improved reproductive tolerance to glyphosate relative to soybean plants comprising the same two transgenes but lacking the dicamba tolerance locus. Such improved reproductive tolerance to glyphosate is reflected in reduced sterility when the plants are exposed to glyphosate.

In certain embodiments, the dicamba tolerance locus provided herein can provide for improved performance of additional combinations of transgenic traits (i.e. “stacked transgenic traits”) in soybean plants. In such embodiments, the dicamba tolerance locus provided herein can be alternatively referred to and considered a “stacked transgenic trait improvement” locus. Allele(s) of the dicamba tolerance locus or “stacked transgenic trait improvement” locus that do not confer such dicamba tolerance or such stacked transgenic trait improvements are referred to herein as dicamba sensitivity or “stacked transgenic trait sensitivity” loci. Transgenic plants comprising the stacked transgenic trait improvement locus provided herein exhibit improved performance of both transgenes present in the transgenic plant relative to plants comprising the same two transgenes that lack the stacked transgenic trait improvement locus. Such improved performance can manifest in any of enhanced transgenic trait performance, increased transgene efficacy, and/or increased transgene expression. Transgenic plants comprising the stacked trait improvement locus and two transgenes are thus provided herein. Thus, in certain embodiments the two independent and distinct transgenes that exhibit improved performance in the presence of the stacked transgenic trait improvement locus both contribute to the same trait. In certain embodiments, this same trait is selected from the group consisting of resistance to a single herbicide, resistance to an insect, resistance to a nematode, resistance to a fungal disease, resistance to an abiotic stress, an improvement in seed oil quantity, an improvement in seed oil quality, and intrinsic yield increases. In certain embodiments, the two transgenes can contribute to the same herbicide resistance trait where the herbicide resistance is selected from the group consisting of glyphosate, dicamba, 2,4-D, glufosinate, bromoxynil, synthetic auxins, and inhibitors of acetolactate synthase (ALS), acetyl CoA carboxylase (ACCase) and hydroxyphenyl pyruvate dioxygenase (HPPD) resistance. In certain embodiments, the two transgenes can contribute to the same insect, fungal, or nematode resistance trait where the resistance to the same insect, fungal, or nematode pest is by a different mode of action to provide for improved pest resistance management. In other embodiments, the two transgenes can be two independent and distinct transgenes that encode different genes but contribute to a different trait. In certain embodiments, this different trait is independently selected from the group consisting of resistance to one or more herbicide(s), resistance to one or more insect(s), resistance to one or more nematode(s), resistance to one or more fungal disease(s), resistance to one or more abiotic stress(es), one or more improvement(s) in seed oil quantity or quantities, one or more improvement(s) in seed oil quality or qualities, intrinsic yield increases, and combinations thereof. In certain embodiments the stacked trait improvement locus is an herbicide tolerance locus that provides for improved tolerance to at least two distinct herbicides in plants comprising at least two transgenes that respectively confer resistance to those two herbicides. In certain embodiments, the herbicide tolerance locus provides for improved tolerance to at least two herbicides selected from the group consisting of glyphosate, dicamba, 2,4-D, glufosinate, bromoxynil, synthetic auxins, and inhibitors of acetolactate synthase (ALS), acetyl CoA carboxylase (ACCase) and hydroxyphenyl pyruvate dioxygenase (HPPD) resistance in plants comprising at least two transgenes that confer resistance to those two herbicides. In certain embodiments, the herbicide tolerance locus confers improved tolerance to dicamba, improved reproductive tolerance to glyphosate, and improved tolerance to a synthetic auxin that includes, but is not limited to 2, 4-D, in a plant comprising transgenes that confer resistance to dicamba, glyphosate, and the synthetic auxin that includes, but is not limited to 2, 4-D. In certain embodiments, the two transgenes can confer a distinct herbicide resistance trait where the herbicide resistance is selected from the group consisting of glyphosate, dicamba, 2,4-D, glufosinate, bromoxynil, synthetic auxins other than 2,4-D, acetolactate synthase (ALS), acetyl CoA carboxylase (ACCase) and hydroxyphenyl pyruvate dioxygenase (HPPD) resistance. Provided herein are soybean plants comprising any combination of a stacked trait improvement locus and at least two transgenes conferring herbicide tolerance selected from the group consisting of glyphosate, dicamba, 2,4-D, glufosinate, bromoxynil, synthetic auxins, and inhibitors of acetolactate synthase (ALS), acetyl CoA carboxylase (ACCase) and hydroxyphenyl pyruvate dioxygenase (HPPD). In certain embodiments, soybean plants comprising an introgressed stacked trait improvement locus, at least one transgene selected from the group consisting of glyphosate, dicamba, 2,4-D, glufosinate, bromoxynil, synthetic auxins other than 2,4-D, acetolactate synthase (ALS), acetyl CoA carboxylase (ACCase), hydroxyphenyl pyruvate dioxygenase (HPPD), and sulfonylurea herbicide resistance transgenes, and at least one transgene selected from the group of transgenes conferring insect resistance, nematode resistance, fungal resistance, an improvement in seed oil quantity, an improvement in seed oil quality, abiotic stress resistance, and intrinsic yield increases are provided. In still other embodiments, soybean plants comprising an introgressed stacked trait improvement locus, at least one transgene selected from the group consisting of glyphosate, dicamba, 2,4-D, and at least one transgene conferring resistance to an insect are provided. In still other embodiments, soybean plants comprising an introgressed stacked trait improvement locus, at least one transgene selected from the group consisting of glyphosate, dicamba, glufosinate, and 2,4-D resistance conferring transgenes, and at least one transgene conferring resistance to an insect that encodes a Bacillus thuringiensis toxin are provided. In certain embodiments, soybean plants comprising an introgressed stacked trait improvement locus, a glyphosate and a dicamba resistance conferring transgene, and a cry1Ac insect resistance transgene are provided. In still other embodiments, soybean plants comprising at least one herbicide resistance transgene selected from the group consisting of a dicamba resistance conferring transgene. a glyphosate resistance conferring transgene, a 2,4-D resistance conferring transgene, and a glufosinate resistance conferring transgene and/or at least one transgene encoding a product that confers insect resistance selected from the group consisting of a dsRNA that inhibits a target gene of an insect pest, a patatin, a Bacillus thuringiensis insecticidal protein, a Xenorhabdus insecticidal protein, a Photorhabdus insecticidal protein, a Bacillus laterosporous insecticidal protein, a Bacillus sphaericus insecticidal protein, and a lignin are provided.

Methods of identifying a soybean plant that comprises a genotype associated with stacked transgenic trait improvement are thus provided. In certain embodiments, the methods can comprise detecting in a soybean plant an allele in at least one genetic locus associated with stacked transgenic trait improvement, where the genetic locus is in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12), and denoting that the plant comprises a genotype associated with stacked transgenic trait improvement. In certain embodiments, the methods can further comprise the step of selecting the denoted plant from a population of plants. In certain embodiments, the plant comprises at least two transgenes that contribute to the same trait. In certain embodiments, this same trait is selected from the group consisting of resistance to a single herbicide, resistance to an insect, resistance to a nematode, resistance to a fungal disease, resistance to an abiotic stress, an improvement in seed oil quantity, an improvement in seed oil quality, and intrinsic yield increases. In certain embodiments, the plant comprises at least two transgenes that contribute to different traits. In certain embodiments, this different trait is independently selected from the group consisting of resistance to one or more herbicide(s), resistance to one or more insect(s), resistance to one or more nematode(s), resistance to one or more fungal disease(s), resistance to one or more abiotic stress(es), one or more improvement(s) in seed oil quantity or quantities, one or more improvement(s) in seed oil quality or qualities, intrinsic yield increases, and combinations thereof. In certain embodiments, the soybean plant or progeny thereof is exposed to a dosage of an herbicide sufficient to cause a deleterious effect in a susceptible variety comprising the transgene conferring resistance to that herbicide but lacking the stacked transgenic trait improvement locus. In certain embodiments of any of the aforementioned methods, a plant that exhibits stacked transgenic trait improvement is selected. In certain embodiments of any of the aforementioned methods, a genotype associated with stacked transgenic trait improvement comprises at least one polymorphic allele of at least one marker in a first sub-region of the linkage group L region that is flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6) and/or at least one polymorphic allele of at least one marker in a second sub-region of the linkage group L region that is flanked by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12) and/or at least one polymorphic allele of at least one marker in a third sub-region of the linkage group L region that is flanked by loci BU55345 (SEQ ID NO:7) and M0114388 (SEQ ID NO:8) is provided. In certain embodiments of any of the aforementioned methods, the genotype associated with stacked transgenic trait improvement comprises at least one polymorphic allele of at least one marker in the linkage group L region selected from the group consisting of a TT allele M0205350 (SEQ ID NO: 10), a TT allele of M0101742 (SEQ ID NO: 5), a CC allele of M0102027 (SEQ ID NO: 11), and an AA allele of NGMAX008197032 (SEQ ID NO:52).

Methods for obtaining a soybean plant comprising in its genome at least one stacked transgenic trait improvement locus are also provided. In certain embodiments, these methods can compromise the steps of: (a) genotyping a plurality of soybean plants with respect to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12); and, (b) selecting a soybean plant comprising in its genome at least one genetic locus comprising a genotype associated with stacked transgenic trait improvement. In certain embodiments, the plant comprises at least two transgenes that contribute to the same trait. In certain embodiments, this same trait is selected from the group consisting of resistance to a single herbicide, resistance to an insect, resistance to a nematode, resistance to a fungal disease, resistance to an abiotic stress, an improvement in seed oil quantity, an improvement in seed oil quality, and intrinsic yield increases. In certain embodiments, the plant comprises at least two transgenes that contribute to different traits. In certain embodiments, this different trait is independently selected from the group consisting of resistance to one or more herbicide(s), resistance to one or more insect(s), resistance to one or more nematode(s), resistance to one or more fungal disease(s), resistance to one or more abiotic stress(es), one or more improvement(s) in seed oil quantity or quantities, one or more improvement(s) in seed oil quality or qualities, intrinsic yield increases, and combinations thereof. In certain embodiments of these methods, the genotype associated with stacked transgenic trait improvement comprises at least one polymorphic allele of at least one marker in a first sub-region of the linkage group L region flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6) and/or at least one polymorphic allele of at least one marker in a second sub-region of the linkage group L region that is flanked by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12) and/or at least one polymorphic allele of at least one marker in a third sub-region of the linkage group L region that is flanked by loci BU55345 (SEQ ID NO:7) and M0114388 (SEQ ID NO:8). In certain embodiments of any of these aforementioned methods, the genotype associated with stacked transgenic trait improvement comprises at least one polymorphic allele of at least one marker in the first linkage group L region, the first sub-region, the second sub-region, the third sub-region, where the marker is selected from the group consisting of a TT allele M0205350 (SEQ ID NO: 10), a TT allele of M0101742 (SEQ ID NO: 5), a CC allele of M0102027 (SEQ ID NO: 11), and an AA allele of NGMAX008197032 (SEQ ID NO:52). In certain embodiments of these methods, the plurality of soybean plants comprises a population that is obtained by: i) crossing a parent plant comprising at least one stacked transgenic trait improvement locus with a parent plant lacking a stacked transgenic trait improvement locus; or, ii) obtaining seed or progeny from a parental plant segregating for at least one stacked transgenic trait improvement locus. In certain embodiments of any of the aforementioned methods, the methods can further comprise the step of assaying for the presence of at least one additional marker, where the additional marker is either linked or unlinked to the linkage group L genomic region. In certain embodiments of any of the aforementioned methods, the plurality of soybean plants, the soybean plant, and/or progeny thereof are exposed to a dosage of an herbicide sufficient to cause a deleterious effect in a susceptible variety comprising the transgene conferring resistance to that herbicide but lacking the stacked transgenic trait improvement locus. In certain embodiments of any of the aforementioned methods, a plant that exhibits stacked transgenic trait improvement is selected.

Methods for producing a soybean plant comprising in its genome at least one introgressed stacked transgenic trait improvement locus are also provided. In certain embodiments, these methods comprise the steps of: (a) crossing a first soybean plant with a stacked transgenic trait improvement locus with a second soybean plant lacking a stacked transgenic trait improvement locus in a first linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6) and/or at least one polymorphic allele of at least one marker in a second sub-region of the linkage group L region that is flanked by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12) and/or at least one polymorphic allele of at least one marker in a third sub-region of the linkage group L region that is flanked by loci BU55345 (SEQ ID NO:7) and M0114388 (SEQ ID NO:8) and at least one linked polymorphic locus not present in the first soybean plant to obtain a population segregating for the stacked transgenic trait improvement loci and the linked polymorphic locus; (b) detecting at least two polymorphic nucleic acids in at least one soybean plant from the population, where at least one of the polymorphic nucleic acids is located in the first linkage group L region and/or the second linkage group L region and where at least one of the polymorphic amino acids is a linked polymorphic locus not present in the first soybean plant; and (c) selecting a soybean plant comprising a genotype associated with stacked transgenic trait improvement and at least one linked marker found in the second soybean plant lacking the stacked transgenic trait improvement locus but not found in the first soybean plant, thereby obtaining a soybean plant comprising in its genome at least one introgressed stacked transgenic trait improvement locus. In certain embodiments, the first and/or second plant comprises at least two transgenes that contribute to the same trait. In certain embodiments, this same trait is selected from the group consisting of resistance to a single herbicide, resistance to an insect, resistance to a nematode, resistance to a fungal disease, resistance to an abiotic stress, an improvement in seed oil quantity, an improvement in seed oil quality, and intrinsic yield increases. In certain embodiments, the plant comprises at least two transgenes that contribute to different traits. In certain embodiments, this different trait is independently selected from the group consisting of resistance to one or more herbicide(s), resistance to one or more insect(s), resistance to one or more nematode(s), resistance to one or more fungal disease(s), resistance to one or more abiotic stress(es), one or more improvement(s) in seed oil quantity or quantities, one or more improvement(s) in seed oil quality or qualities, intrinsic yield increases, and combinations thereof. In certain embodiments of these methods, the population, the selected soybean plant, and/or progeny of selected soybean plant is exposed to a dosage of an herbicide sufficient to cause a deleterious effect in a susceptible variety comprising the transgene that confers resistance to the herbicide but lacking the stacked transgenic trait improvement locus. In certain embodiments of these methods, the polymorphic nucleic acid detected in step (b) is detected with at least one marker selected from the group consisting of M0205350 (SEQ ID NO: 10), M0101742 (SEQ ID NO: 5), M0102027 (SEQ ID NO: 11), and an AA allele of NGMAX008197032 (SEQ ID NO:52). In certain embodiments of these methods, the polymorphic nucleic acid detected in step (b) comprises a TT allele of M0205350 (SEQ ID NO: 10), a TT allele of M0101742 (SEQ ID NO: 5), a CC allele of M0102027 (SEQ ID NO: 11) and an AA allele of NGMAX008197032 (SEQ ID NO:52). In certain embodiments of these methods, the polymorphic nucleic acid detected in step (b) is detected with marker M0205350 (SEQ ID NO: 10), M0102027 (SEQ ID NO: 11) or NGMAX008197032 (SEQ ID NO: 52). In certain embodiments of these methods, the polymorphic nucleic acids are detected with marker M0101742 (SEQ ID NO: 5). In certain embodiments of these methods, the polymorphic nucleic acids are detected with marker NGMAX008197032 (SEQ ID NO:52). In certain embodiments of any of the aforementioned methods, the linked polymorphic locus is detected with a genotypic marker, a phenotypic marker, or both. In certain embodiments of these methods, the linked polymorphic locus is detected with a marker that is located within about 1000, 500, 100, 40, 20, 10, or 5 kilobases (Kb) of the stacked transgenic trait improvement locus. In certain embodiments of these methods, the linked polymorphic locus is detected with at least one marker selected from the group consisting of asmbl_11856 (SEQ ID NO: 1), TC122822 (SEQ ID NO: 2), BI967232 (SEQ ID NO: 3), M0205537 (SEQ ID NO: 15), M0202715 (SEQ ID NO: 16), M0206286 (SEQ ID NO: 17), M0206054 (SEQ ID NO: 18), and M0205375 (SEQ ID NO: 19).

Transgenic soybean plants comprising introgressed linkage group L regions comprising at least one polymorphic allele of at least one marker in a first sub-region of the linkage group L region that flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6) and/or at least one polymorphic allele of at least one marker in a second sub-region of the linkage group L region that is flanked by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12) and/or at least one polymorphic allele of at least one marker in a third sub-region of the linkage group L region that is flanked by loci BU55345 (SEQ ID NO:7) and M0114388 (SEQ ID NO:8), where the polymorphic alleles are associated with a stacked transgenic trait improvement locus, and where the plant comprises: (a) at least two transgenes that contribute to the same trait; or, (b) at least two transgenes that contribute different traits. In certain embodiments, this same trait is selected from the group consisting of resistance to a single herbicide, resistance to an insect, resistance to a nematode, resistance to a fungal disease, resistance to an abiotic stress, an improvement in seed oil quantity, an improvement in seed oil quality, and intrinsic yield increases. In certain embodiments, this different trait is independently selected from the group consisting of resistance to one or more herbicide(s), resistance to one or more insect(s), resistance to one or more nematode(s), resistance to one or more fungal disease(s), resistance to one or more abiotic stress(es), one or more improvement(s) in seed oil quantity or quantities, one or more improvement(s) in seed oil quality or qualities, intrinsic yield increases, and combinations thereof. In certain embodiments, the polymorphic alleles comprise a TT allele of M0205350 (SEQ ID NO: 10), a TT allele of M0101742 (SEQ ID NO: 5), a CC allele of M0102027 (SEQ ID NO: 11), and an AA allele of NGMAX008197032 (SEQ ID NO:52). In certain embodiments, the transgenic plant exhibits a stacked transgenic trait improvement. In certain embodiments, the transgenic plant further comprises a transgene that confers resistance to glyphosate and exhibits reproductive tolerance to glyphosate. In certain embodiments of any one of the aforementioned methods, the plant further comprises at least one of a 2,4-D, glufosinate, bromoxynil, synthetic auxins other than 2,4-D, acetolactate synthase (ALS), acetyl CoA carboxylase (ACCase), hydroxyphenyl pyruvate dioxygenase (HPPD), or sulfonylurea herbicide resistance transgenes and/or at least one transgene selected from the group of transgenes conferring insect resistance, nematode resistance, fungal resistance, an improvement in seed oil quantity, an improvement in seed oil quality, abiotic stress resistance, and intrinsic yield increases.

Also provided herein are methods of identifying a soybean plant that comprises a genotype associated with stacked transgenic trait improvement, comprising: detecting in a soybean plant an allele in at least one genetic locus associated with stacked transgenic trait improvement, wherein the genetic locus is in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12), and denoting that the plant comprises a genotype associated with stacked transgenic trait improvement. In certain embodiments, the method further comprises the step of selecting the denoted plant from a population of plants and wherein the detection is performed either before or after the selection. In certain embodiments, the denoted plant comprises at least one transgene that confer resistance to an herbicide and is selected for improved tolerance to that herbicide. In certain embodiments, the selection comprises exposing the population of plants to a dosage of herbicide sufficient to cause a deleterious effect in a susceptible variety comprising the transgene that confers resistance to the herbicide. In certain embodiments: (i) the plants comprise an herbicide resistance transgene selected from the group consisting of a dicamba resistance conferring transgene, a glyphosate resistance conferring transgene, a 2,4-D resistance conferring transgene, and a glufosinate resistance conferring transgene; and (ii) the plants comprising the herbicide resistance transgene are exposed to a dosage of a corresponding herbicide selected from the group consisting of dicamba, glyphosate, glufosinate, and 2,4-D that is sufficient to cause a deleterious effect in a susceptible variety comprising the herbicide resistant transgene that confers resistance to the corresponding herbicide. In certain embodiments, the stacked transgenic trait improvement is independently selected from the group consisting of an improvement in transgene-mediated resistance to one or more herbicide(s), an improvement in transgene-mediated resistance to one or more insect(s), an improvement in transgene-mediated resistance to one or more nematode(s), an improvement in transgene-mediated resistance to one or more fungal disease(s), an improvement in transgene-mediated resistance to one or more abiotic stress(es), in one or more improvement(s) in transgene-mediated seed oil quantity trait(s), one or more improvement(s) in seed oil quality trait(s), an improvement in transgene-mediated intrinsic yield increases, and combinations thereof. In certain embodiments, the denoted plant comprises at least one herbicide resistance transgene and/or at least one insect resistance conferring transgene that encodes a Bacillus thuringiensis toxin. In certain embodiments, the denoted plant comprises at least one herbicide resistance transgene selected from the group consisting of a dicamba resistance conferring transgene. a glyphosate resistance conferring transgene, a 2,4-D resistance conferring transgene, and a glufosinate resistance conferring transgene and/or at least one transgene encoding a product that confers insect resistance selected from the group consisting of a dsRNA that inhibits a target gene of an insect pest, a patatin, a Bacillus thuringiensis insecticidal protein, a Xenorhabdus insecticidal protein, a Photorhabdus insecticidal protein, a Bacillus laterosporous insecticidal protein, a Bacillus sphaericus insecticidal protein, and a lignin. In certain embodiments of any of the preceding methods, the genotype associated with stacked transgenic trait improvement comprises at least one polymorphic allele of at least one marker in a first sub-region of the linkage group L region that is flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6) and/or at least one polymorphic allele of at least one marker in a second sub-region of the linkage group L region that is flanked by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12) and/or at least one polymorphic allele of at least one marker in a third sub-region of the linkage group L region that is flanked by loci BU55345 (SEQ ID NO:7) and M0114388 (SEQ ID NO:8). In certain embodiments of any of the preceding methods, the genotype associated with stacked transgenic trait improvement comprises at least one polymorphic allele of at least one marker in the linkage group L region selected from the group consisting of a TT allele M0205350 (SEQ ID NO: 10), a TT allele of M0101742 (SEQ ID NO: 5), a CC allele of M0102027 (SEQ ID NO: 11), and an AA allele of NGMAX008197032 (SEQ ID NO:52).

Also provided are methods for obtaining a soybean plant comprising in its genome at least one stacked transgenic trait improvement locus, compromising the steps of: genotyping a plurality of soybean plants with respect to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12); and selecting a soybean plant comprising in its genome at least one genetic locus comprising a genotype associated with stacked transgenic trait improvement. In certain embodiments of the methods, the genotype associated with stacked transgenic trait improvement comprises at least one polymorphic allele of at least one marker in a first sub-region of the linkage group L region flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6) and/or at least one polymorphic allele of at least one marker in a second sub-region of the linkage group L region that is flanked by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12) and/or at least one polymorphic allele of at least one marker in a third sub-region of the linkage group L region that is flanked by loci BU55345 (SEQ ID NO:7) and M0114388 (SEQ ID NO:8). In certain embodiments of any of the preceding methods, the genotype associated with stacked transgenic trait improvement comprises at least one polymorphic allele of at least one marker in the first linkage group L region, the first sub-region, or the second sub-region, wherein the marker is selected from the group consisting of a TT allele M0205350 (SEQ ID NO: 10), a TT allele of M0101742 (SEQ ID NO: 5), a CC allele of M0102027 (SEQ ID NO: 11), and an AA allele of NGMAX008197032 (SEQ ID NO:52). In certain embodiments of the methods, the plurality of soybean plants comprises a population that is obtained by: i) crossing a parent plant comprising at least one stacked transgenic trait improvement locus with a parent plant comprising at least one stacked transgenic trait sensitivity locus; or, ii) obtaining seed or progeny from a parental plant segregating for at least one stacked transgenic trait improvement locus. In certain embodiments of the methods, the population contains plants that comprise at least one transgene that confers resistance to an herbicide and the stacked transgenic trait improvement comprises improved tolerance to a corresponding herbicide. In certain embodiments of any of the preceding methods, the methods further comprise the step of assaying for the presence of at least one additional marker, wherein the additional marker is either linked or unlinked to the linkage group L genomic region. In certain embodiments of any of the preceding methods, the plurality of soybean plants, the soybean plant, and/or progeny thereof are exposed to a dosage of herbicide sufficient to cause a deleterious effect in a susceptible variety comprising the transgene that confers resistance to the herbicide. In certain embodiments of any of the preceding methods, a plant that exhibits dicamba tolerance and/or reproductive tolerance to glyphosate and/or glufosinate tolerance and/or 2,4-D tolerance is selected. In certain embodiments of any of the preceding methods, the stacked transgenic trait improvement is selected from the group consisting of an improvement in transgene-mediated resistance to one or more herbicide(s), an improvement in transgene-mediated resistance to one or more insect(s), an improvement in transgene-mediated resistance to one or more nematode(s), an improvement in transgene-mediated resistance to one or more fungal disease(s), an improvement in transgene-mediated resistance to one or more abiotic stress(es), in one or more improvement(s) in transgene-mediated seed oil quantity trait(s), one or more improvement(s) in seed oil quality trait(s), an improvement in transgene-mediated intrinsic yield increases, and combinations thereof.

Also provided herein are methods for producing a soybean plant comprising in its genome at least one introgressed stacked transgenic trait improvement locus comprising the steps of: crossing a first soybean plant with a stacked transgenic trait improvement locus with a second soybean plant comprising: a stacked transgenic trait sensitivity locus in a first linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6) and/or at least one polymorphic allele of at least one marker in a second sub-region of the linkage group L region that is flanked by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12) and/or at least one polymorphic allele of at least one marker in a third sub-region of the linkage group L region that is flanked by loci BU55345 (SEQ ID NO:7) and M0114388 (SEQ ID NO:8) and at least one linked polymorphic locus not present in the first soybean plant to obtain a population segregating for the stacked transgenic trait improvement loci and the linked polymorphic locus; detecting at least two polymorphic nucleic acids in at least one soybean plant from the population, wherein at least one of the polymorphic nucleic acids is located in the first linkage group L region and/or the second linkage group L region and at least one of the polymorphic amino acids is a linked polymorphic locus not present in the first soybean plant; and selecting a soybean plant comprising a genotype associated with stacked transgenic trait improvement and at least one linked marker found in the second soybean plant comprising a stacked transgenic trait sensitivity locus but not in the first soybean plant, thereby obtaining a soybean plant comprising in its genome at least one introgressed stacked transgenic trait improvement locus. In certain embodiments of the methods, at least one of the first or the second soybean plants comprises a transgene that confers resistance to an herbicide. In certain embodiments of the methods, the population, the selected soybean plant, and/or progeny of selected soybean plant is exposed to a dosage of herbicide sufficient to cause a deleterious effect in a susceptible variety comprising the transgene that confers resistance to a corresponding herbicide. In certain embodiments of the methods, the polymorphic nucleic acid detected in step (b) is detected with at least one marker selected from the group consisting of M0205350 (SEQ ID NO: 10), M0101742 (SEQ ID NO: 5), M0102027 (SEQ ID NO: 11), and NGMAX008197032 (SEQ ID NO:52). In certain embodiments of the methods, the polymorphic nucleic acid detected in step (b) comprises a TT allele of M0205350 (SEQ ID NO: 10), a TT allele of M0101742 (SEQ ID NO: 5), a CC allele of M0102027 (SEQ ID NO: 11), and an AA allele of NGMAX008197032 (SEQ ID NO:52). In certain embodiments of the methods, the polymorphic nucleic acid detected in step (b) is detected with marker M0205350 (SEQ ID NO: 10), M0102027 (SEQ ID NO: 11), or marker NGMAX008197032 (SEQ ID NO:52). In certain embodiments of the methods, the polymorphic nucleic acids are detected with marker M0101742 (SEQ ID NO: 5). In certain embodiments of any of the preceding methods, the linked polymorphic locus is detected with a genotypic marker, a phenotypic marker, or both. In certain embodiments, the linked polymorphic locus is detected with a marker that is located within about 1000, 500, 100, 40, 20, 10, or 5 kilobases (Kb) of the stacked transgenic trait improvement locus. In certain embodiments, the linked polymorphic locus is detected with at least one marker selected from the group consisting of asmbl_11856 (SEQ ID NO: 1), TC122822 (SEQ ID NO: 2), BI967232 (SEQ ID NO: 3), M0205537 (SEQ ID NO: 15), M0202715 (SEQ ID NO: 16), M0206286 (SEQ ID NO: 17), M0206054 (SEQ ID NO:18), and M0205375 (SEQ ID NO: 19).

Also provided are transgenic soybean plants comprising introgressed linkage group L regions comprising at least one polymorphic allele of at least one marker in a first sub-region of the linkage group L region that flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6) and/or at least one polymorphic allele of at least one marker in a second sub-region of the linkage group L region that is flanked by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12) and/or at least one polymorphic allele of at least one marker in a third sub-region of the linkage group L region that is flanked by loci BU55345 (SEQ ID NO:7) and M0114388 (SEQ ID NO:8), wherein the polymorphic alleles are associated with stacked transgenic trait improvement and wherein the plant comprises at least one transgene. In certain embodiments, the transgene confers resistance to an herbicide. In certain embodiments, the polymorphic alleles comprise a TT allele of M0205350 (SEQ ID NO: 10), a TT allele of M0101742 (SEQ ID NO: 5), a CC allele of M0102027 (SEQ ID NO: 11), and an AA allele of NGMAX008197032 (SEQ ID NO:52). In certain embodiments of any of the preceding methods, the plant exhibits tolerance to at least one herbicide. In certain embodiments, the plant comprises: i) a transgene that confers resistance to glyphosate and exhibits reproductive tolerance to glyphosate; and/or (ii) a dicamba resistance conferring transgene and exhibits dicamba tolerance; and/or (iii) a glufosinate resistance conferring transgene and exhibits glufosinate tolerance; and/or (iv) a 2,4-D resistance conferring transgene and exhibits 2,4-D tolerance. In certain embodiments, the plant comprises at least one transgene conferring resistance to a herbicide selected from the group consisting of dicamba, 2,4-D, glufosinate, bromoxynil, synthetic auxins other than 2,4-D, acetolactate synthase (ALS), acetyl CoA carboxylase (ACCase), hydroxyphenyl pyruvate dioxygenase (HPPD), and a sulfonylurea herbicide and/or at least one transgene selected from the group of transgenes conferring insect resistance, nematode resistance, fungal resistance, an improvement in seed oil quantity, an improvement in seed oil quality, abiotic stress resistance, and intrinsic yield increases. In certain embodiments, the plant comprises at least one herbicide resistance transgene and/or at least one insect resistance conferring transgene that encodes a Bacillus thuringiensis toxin. In certain embodiments, the plant comprises at least one herbicide resistance transgene selected from the group consisting of a dicamba resistance conferring transgene a glyphosate resistance conferring transgene, a 2,4-D resistance conferring transgene, and a glufosinate resistance conferring transgene and/or at least one transgene encoding a product that confers insect resistance selected from the group consisting of a dsRNA that inhibits a target gene of an insect pest, a patatin, a Bacillus thuringiensis insecticidal protein, a Xenorhabdus insecticidal protein, a Photorhabdus insecticidal protein, a Bacillus laterosporous insecticidal protein, a Bacillus sphaericus insecticidal protein, and a lignin.

Also provided herein are methods of identifying a transgenic soybean plant that comprises a genotype associated with stacked transgenic trait improvement, the method comprising: (a) scoring at least one transgenic plant in a population of transgenic soybean plants that had been exposed to dicamba for dicamba tolerance, the plants having a transgene that confers resistance to dicamba; and, (b) selecting a transgenic plant that exhibits dicamba tolerance, thereby identifying a transgenic soybean plant that comprises a genotype associated with stacked transgenic trait improvement. In certain embodiments, the population is segregating for to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12) that is associated with stacked transgenic trait improvement. In certain embodiments, the method further comprises genotyping the selected soybean plant with respect to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12). In certain embodiments, the selected transgenic plant further comprises a transgene that confers resistance to glyphosate and the selected transgenic plant or progeny thereof is scored for reproductive tolerance to glyphosate following exposure to glyphosate. In certain embodiments of any of the preceding methods, the methods further comprise exposing the population of transgenic soybean plants to dicamba. In certain embodiments of any of the preceding methods, dicamba tolerance is scored by determining a reduction in malformation when compared to a dicamba sensitive transgenic plant that comprises the transgene that confers resistance to dicamba. In certain embodiments of any of the preceding methods, the stacked transgenic trait improvement is selected from the group consisting of an improvement in transgene-mediated resistance to one or more herbicide(s), an improvement in transgene-mediated resistance to one or more insect(s), an improvement in transgene-mediated resistance to one or more nematode(s), an improvement in transgene-mediated resistance to one or more fungal disease(s), an improvement in transgene-mediated resistance to one or more abiotic stress(es), in one or more improvement(s) in transgene-mediated seed oil quantity trait(s), one or more improvement(s) in seed oil quality trait(s), an improvement in transgene-mediated intrinsic yield increases, and combinations thereof. In certain embodiments of any of the preceding methods, the selected plant comprises at least one additional herbicide resistance transgene selected from the group consisting of a glyphosate resistance conferring transgene, a 2,4-D resistance conferring transgene, and a glufosinate resistance conferring transgene and/or at least transgene encoding a product that confers insect resistance selected from the group consisting of a dsRNA that inhibits a target gene of an insect pest, a patatin, a Bacillus thuringiensis insecticidal protein, a Xenorhabdus insecticidal protein, a Photorhabdus insecticidal protein, a Bacillus laterosporous insecticidal protein, a Bacillus sphaericus insecticidal protein, and a lignin.

Also provided herein are methods of identifying a transgenic soybean plant that comprises a genotype associated with stacked transgenic trait improvement, comprising: (a) scoring at least one plant in a population of transgenic soybean plants that had been exposed to glyphosate for reproductive tolerance to glyphosate, wherein the plants comprise a transgene that confers resistance to glyphosate; and, (b) selecting a transgenic plant that exhibits reproductive tolerance to glyphosate, thereby identifying a transgenic soybean plant that comprises a genotype associated with stacked transgenic trait improvement. In certain embodiments, the population is segregating for to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12) that is associated with stacked transgenic trait improvement. In certain embodiments, the method further comprises genotyping the selected soybean plant with respect to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12). In certain embodiments of any of the preceding methods, the selected transgenic plant further comprises a transgene that confers resistance to dicamba and the selected transgenic plant or progeny thereof is scored for tolerance to dicamba following exposure to dicamba. In certain embodiments of any of the preceding methods, the methods further comprise exposing the population of transgenic soybean plants to glyphosate. In certain embodiments of any of the preceding methods, glyphosate reproductive tolerance is scored by determining a reduction in sterility when compared to a transgenic plant that exhibits glyphosate reproductive sensitivity and comprises the transgene that confers resistance to glyphosate. In certain embodiments of any of the preceding methods, the stacked transgenic trait improvement is selected from the group consisting of an improvement in transgene-mediated resistance to one or more herbicide(s), an improvement in transgene-mediated resistance to one or more insect(s), an improvement in transgene-mediated resistance to one or more nematode(s), an improvement in transgene-mediated resistance to one or more fungal disease(s), an improvement in transgene-mediated resistance to one or more abiotic stress(es), in one or more improvement(s) in transgene-mediated seed oil quantity trait(s), one or more improvement(s) in seed oil quality trait(s), an improvement in transgene-mediated intrinsic yield increases, and combinations thereof. In certain embodiments of any of the preceding methods, the selected plant comprises at least one additional herbicide resistance transgene selected from the group consisting of a dicamba resistance conferring transgene, a 2,4-D resistance conferring transgene, and a glufosinate resistance conferring transgene and/or at least one transgene encoding a product that confers insect resistance selected from the group consisting of a dsRNA that inhibits a target gene of an insect pest, a patatin, a Bacillus thuringiensis insecticidal protein, a Xenorhabdus insecticidal protein, a Photorhabdus insecticidal protein, a Bacillus laterosporous insecticidal protein, a Bacillus sphaericus insecticidal protein, and a lignin.

Also provided are methods of obtaining a transgenic soybean plant that comprises a genotype associated with stacked transgenic trait improvement, the methods comprising: exposing a population of transgenic soybean plants to an herbicide, wherein the plants have a transgene that confers resistance to the herbicide; observing herbicide tolerance exhibited by one or more soybean plants following exposure to the herbicide; and, (c) selecting a transgenic plant that exhibits herbicide tolerance, thereby obtaining a transgenic soybean plant that comprises a genotype associated with stacked transgenic trait improvement. In certain embodiments, the population is segregating for to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12) that is associated with stacked transgenic trait improvement. In certain embodiments, the method further comprises genotyping the selected soybean plant with respect to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12). In certain embodiments of any of the preceding methods, the transgene that confers resistance to the herbicide is selected from the group consisting of a dicamba resistance conferring transgene, a glyphosate resistance conferring transgene, a 2,4-D resistance conferring transgene, and a glufosinate resistance conferring transgene and the plants are exposed to the corresponding herbicide. In certain embodiments of any of the preceding methods, the transgene confers resistance to glyphosate and the selected transgenic plant or progeny thereof is scored for reproductive tolerance to glyphosate following exposure to glyphosate. In certain embodiments of any of the preceeding methods, the transgene confers resistance to dicamba and the selected transgenic plant or progeny thereof are scored for dicamba tolerance.

Also provided herein are methods of identifying a transgenic soybean plant that comprises a genotype associated with reproductive tolerance to glyphosate, the method comprising: (a) scoring at least one transgenic plant in a population of transgenic soybean plants that had been exposed to dicamba for dicamba tolerance, the plants having a transgene that confers resistance to dicamba; and, (b) selecting a transgenic plant that exhibits dicamba tolerance, thereby identifying a transgenic soybean plant that comprises a genotype associated with reproductive tolerance to glyphosate. In certain embodiments of the methods, the population is segregating for to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12) that is associated with dicamba tolerance. In certain embodiments of the methods, the methods further comprise genotyping the selected soybean plant with respect to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12). In certain embodiments of any of the aforementioned methods, the selected transgenic plant further comprises a transgene that confers resistance to glyphosate and wherein the selected transgenic plant or progeny thereof is scored for reproductive tolerance to glyphosate following exposure to glyphosate. In certain embodiments of any of the aforementioned methods, the methods further comprise exposing the population of transgenic soybean plants to dicamba. In certain embodiments of any of the aforementioned methods, the dicamba tolerance is scored by determining a reduction in malformation when compared to a dicamba sensitive transgenic plant that comprises the transgene that confers resistance to dicamba.

Also provided herein are methods of identifying a transgenic soybean plant that comprises a genotype associated with tolerance to dicamba, comprising: (a) scoring at least one plant in a population of transgenic soybean plants that had been exposed to glyphosate for reproductive tolerance to glyphosate, wherein the plants comprise a transgene that confers resistance to glyphosate; and, (b) selecting a transgenic plant that exhibits reproductive tolerance to glyphosate, thereby identifying a transgenic soybean plant that comprises a genotype associated with dicamba tolerance. In certain embodiments of the methods, the population is segregating for to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12) that is associated with dicamba tolerance. In certain embodiments of the methods, the method further comprises genotyping the selected soybean plant with respect to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12). In certain embodiments of any of the aforementioned methods, the selected transgenic plant further comprises a transgene that confers resistance to dicamba and wherein the selected transgenic plant or progeny thereof is scored for tolerance to dicamba following exposure to dicamba. In certain embodiments of any of the aforementioned methods, the methods further comprise exposing the population of transgenic soybean plants to glyphosate. In certain embodiments of any of the aforementioned methods, the glyphosate reproductive tolerance is scored by determining a reduction in sterility when compared to a transgenic plant that exhibits glyphosate reproductive sensitivity and comprises the transgene that confers resistance to glyphosate.

Also provided herein are methods of obtaining a transgenic soybean plant that comprises a genotype associated with reproductive tolerance to glyphosate, the methods comprising: (a) exposing a population of transgenic soybean plants to dicamba, wherein the plants have a transgene that confers resistance to dicamba; (b) observing dicamba tolerance exhibited by one or more soybean plants following exposure to dicamba; and, (c) selecting a transgenic plant that exhibits dicamba tolerance, thereby obtaining a transgenic soybean plant that comprises a genotype associated with reproductive tolerance to glyphosate. In certain embodiments of the methods, the population is segregating for to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12) that is associated with dicamba tolerance. In certain embodiments of the methods, the methods further comprise genotyping the selected soybean plant with respect to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12). In certain embodiments of any of the aforementioned methods, the selected transgenic plant further comprises a transgene that confers resistance to glyphosate and the selected transgenic plant or progeny thereof is scored for reproductive tolerance to glyphosate following exposure to glyphosate. In certain embodiments of any of the aforementioned methods, the dicamba tolerance is scored by determining a reduction in malformation when compared to a dicamba sensitive transgenic plant that comprises the transgene that confers resistance to dicamba. Further areas of applicability will become apparent from the description provided herein. It should be understood that the description and specific examples are intended for purposes of illustration only and are not intended to limit the scope of the present disclosure.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a bar graph of the percent injury at 7 days after treatment with Dicamba (y-axis) for various favorable and unfavorable soybean plant haplotypes containing a dicamba resistance conferring transgene (x-axis). The favorable haplotypes are haplotypes that are not associated with the dicamba intolerance trait and the unfavorable haplotypes are associated with the dicamba intolerance trait. In the graph, “Fav Hap is “Favorable Haplotype” and “Unf Hap” is “Unfavorable Haplotype”. The data show that presence of the favorable haplotype is associated with improved tolerance to dicamba.

FIG. 2 shows a bar graph of the percent injury at 7 days after treatment with either a single treatment (V6 stage, solid bars) or two treatments (V3+V6 stages, open bars) of Dicamba (y-axis) for various favorable and unfavorable soybean plant haplotypes (x-axis) containing a dicamba resistance conferring transgene (x-axis). In the graph, “Fav Hap is “Favorable Haplotype” and “Unf Hap” is “Unfavorable Haplotype”. The data show that favorable haplotypes can be selected with either a single treatment at the V6 stage or two treatments at the V3 and V6 stages.

DETAILED DESCRIPTION Definitions

As used herein, an “allele” refers to one of two or more alternative forms of a genomic sequence at a given locus on a chromosome. When all the alleles present at a given locus on a chromosome are the same, that plant is homozygous at that locus. If the alleles present at a given locus on a chromosome differ, that plant is heterozygous at that locus.

As used herein, the term “denoting” when used in reference to a plant genotype refers to any method whereby a plant is indicated to have a certain genotype. Such indications of a certain genotype include, but are not limited to, any method where a plant is physically marked or tagged. Physical markings or tags that can be used include, but not limited to, a barcode, a radio-frequency identification (RFID), a label or the like. Indications of a certain genotype also include, but are not limited to, any entry into any type of written or electronic database whereby the plant's genotype is provided.

A “locus” is a position on a genomic sequence that is usually found by a point of reference; e.g., a short DNA sequence that is a gene, or part of a gene or intergenic region. A locus may refer to a nucleotide position at a reference point on a chromosome, such as a position from the end of the chromosome.

As used herein, “linkage group L” corresponds to the soybean linkage group L described in Choi, et al., Genetics. 2007 May; 176(1): 685-696. Linkage group L, as used herein, also corresponds to soybean chromosome 19 (as described on the World Wide Web at soybase.org/LG2Xsome.php). As used herein, “polymorphism” means the presence of one or more variations of a nucleic acid sequence at one or more loci in a population of at least two members. The variation can comprise but is not limited to one or more nucleotide base substitutions, the insertion of one or more nucleotides, a nucleotide sequence inversion, and/or the deletion of one or more nucleotides.

As used herein, the term “single nucleotide polymorphism,” also referred to by the abbreviation “SNP,” means a polymorphism at a single site wherein the polymorphism constitutes any or all of a single base pair change, an insertion of one or more base pairs, and/or a deletion of one or more base pairs.

As used herein, “marker” means a detectable characteristic that can be used to discriminate between organisms. Examples of such characteristics include, but are not limited to, genetic markers, biochemical markers, fermentation yield, fermentation efficiency, energy yield, secondary compounds, metabolites, morphological characteristics, and agronomic characteristics.

As used herein, “marker assay” means a method for detecting a polymorphism at a particular locus using a particular method. Marker assays thus include, but are not limited to, measurement of at least one phenotype (such as seed color, flower color, or other visually detectable trait as well as any biochemical trait), restriction fragment length polymorphism (RFLP), single base extension, electrophoresis, sequence alignment, allelic specific oligonucleotide hybridization (ASO), random amplified polymorphic DNA (RAPD), microarray-based polymorphism detection technologies, and the like.

As used herein, “genotype” means the genetic component of the phenotype and it can be indirectly characterized using markers or directly characterized by nucleic acid sequencing.

As used herein, the term “introgressed”, when used in reference to a genetic locus, refers to a genetic locus that has been introduced into a new genetic background. Introgression of a genetic locus can thus be achieved through both plant breeding methods or by molecular genetic methods. Such molecular genetic methods include, but are not limited to, various plant transformation techniques and/or methods that provide for homologous recombination, non-homologous recombination, site-specific recombination, and/or genomic modifications that provide for locus substitution or locus conversion. In certain embodiments, introgression could thus be achieved by substitution of a dicamba intolerance locus with a corresponding dicamba tolerance locus or by conversion of a locus from a dicamba intolerance genotype to a dicamba tolerance genotype.

As used herein, “phenotype” means the detectable characteristics of a cell or organism which can be influenced by gene expression.

As used herein, “linkage” refers to relative frequency at which types of gametes are produced in a cross. For example, if locus A has genes “A” or “a” and locus B has genes “B” or “b” and a cross between parent I with AABB and parent B with aabb will produce four possible gametes where the genes are segregated into AB, Ab, aB and ab. The null expectation is that there will be independent equal segregation into each of the four possible genotypes, i.e. with no linkage ¼ of the gametes will of each genotype. Segregation of gametes into a genotypes differing from ¼ are attributed to linkage.

As used herein, the termed “linked”, when used in the context of markers and/or genomic regions, means that the markers and/or genomic regions are located on the same linkage group or chromosome.

As used herein, a “nucleic acid molecule,” be it a naturally occurring molecule or otherwise may be “substantially purified”, if desired, referring to a molecule separated from substantially all other molecules normally associated with it in its native state. More preferably, a substantially purified molecule is the predominant species present in a preparation. A substantially purified molecule may be at least about 60% free, preferably at least about 75% free, more preferably at least about 90% free, and most preferably at least about 95% free from the other molecules (exclusive of solvent) present in the natural mixture. The term “substantially purified” is not intended to encompass molecules present in their native state.

As used herein, “quantitative trait locus (QTL)” means a locus that controls to some degree numerically representable traits that are usually continuously distributed. As used herein, the term “transgene” means nucleic acid molecules in the form of DNA, such as cDNA or genomic DNA, and RNA, such as mRNA or microRNA, which may be single or double stranded.

As used herein, the term “event”, when used in the context of describing a transgenic plant, refers to a particular transformed plant line. In a typical transgenic breeding program, a transformation construct responsible for a trait is introduced into the genome via a transformation method. Numerous independent transformants (events) are usually generated for each construct. These events are evaluated to select those with superior performance.

As used herein, the term “soybean” means Glycine max and includes all plant varieties that can be bred with soybean, including wild soybean species. In certain embodiments, soybean plants from the species Glycine max and the subspecies Glycine max L. ssp. max or Glycine max ssp. formosana can be genotyped using the compositions and methods of the present invention. In an additional aspect, the soybean plant is from the species Glycine soja, otherwise known as wild soybean, can be genotyped using these compositions and methods. Alternatively, soybean germplasm derived from any of Glycine max, Glycine max L. ssp. max, Glycine max ssp. Formosana, and/or Glycine soja can be genotyped using compositions and methods provided herein.

As used herein, the term “bulk” refers to a method of managing a segregating population during inbreeding that involves growing the population in a bulk plot, harvesting the self-pollinated seed of plants in bulk, and using a sample of the bulk to plant the next generation.

As used herein, the phrase “transgene that confers tolerance to dicamba” refers to the ability of a transgene to provide a soybean plant capable of surviving exposure to dicamba at a rate of about 0.5 pounds of acid equivalent per acre of dicamba acid to about 1.5 pounds of acid equivalent per acre of dicamba acid applied at either pre-emergence and/or postemergence. Transgenic plants comprising a transgene that confers tolerance to dicamba can exhibit either a “dicamba tolerant” phenotype in certain soybean germplasms or a “dicamba sensitive” phenotype in other distinct soybean germplasms when exposed to dicamba.

As used herein, the phrase “dicamba intolerant” refers to undesirable phenotypic traits observed in certain soybean germplasms that comprise a transgene that confers resistance to dicamba after exposure to dicamba at a rate of about 0.5 pounds of acid equivalent per acre of dicamba acid to about 1.5 pounds of acid equivalent per acre of dicamba acid. Such undesirable phenotypic traits include, but are not limited to, pronounced bending/twisting of the main stem and petioles, necrosis of the upper nodes and petioles, and/or limitation of new growth.

As used herein, the phrase “dicamba tolerant” refers to either the absence or reduction of undesirable phenotypic traits observed after exposure to dicamba in “dicamba intolerant” soybean germplasms that comprise a transgene that confers resistance to dicamba.

As used herein, the term “comprising” means “including but not limited to”.

As used herein, the terms “scoring” or “score”, refer to any qualitive, semi-quantitive, or quantitive method for determining the presence, absence, and/or the partial presence or absence, of a phenotypic trait.

As used herein, the phrase “susceptible variety”, when used in reference to herbicide tolerance in a soybean plant comprising a transgene that confers resistance to that herbicide, refers to a soybean variety that allele(s) of the stacked transgenic trait improvement locus that do not confer such stacked transgenic trait improvements. “Susceptible varieties’ are also referred to herein as “sensitive varieties” in the context of herbicide tolerance in a soybean plant comprising a transgene that confers resistance to that herbicide.

As used herein, the phrase “corresponding herbicide”, when used in reference to a transgene that confers herbicide resistance, refers to the herbicide that the transgene confers resistance to. Thus, a corresponding herbicide for a transgene that confers resistance to glyphosate, dicamba, 2,4-D, or glufosinate is respectively glyphosate, dicamba, 2,4-D, or glufosinate.

DESCRIPTION

In accordance with the present invention, Applicants have discovered genomic regions, associated markers, and associated methods for identifying and associating genotypes that effect the levels of dicamba tolerance observed in soybean plants comprising a transgene that confers resistance to dicamba. Dicamba (3, 6-dichloro-o-anisic acid) is a useful broad spectrum herbicide for controlling weeds. For example, in one embodiment, a method of the invention comprises screening a plurality of transgenic germplasm entries displaying a heritable variation for at least one transgene mediated dicamba resistance trait wherein the heritable variation is linked to at least one genotype; and associating at least one genotype from the transgenic germplasm entries to at least one dicamba tolerance trait. In another embodiment, a method of the invention comprises crossing at least two germplasm entries with a test germplasm entry for the evaluation of performance of at least one dicamba tolerance trait in order to determine preferred crossing schemes. The methods of the present invention can be used with traditional breeding techniques as described below to more efficiently screen and identify genotypes affecting a dicamba tolerance trait.

The use of markers to infer a phenotype of interest results in the economization of a breeding program by substituting costly, time-intensive phenotyping assays with genotyping assays. Further, breeding programs can be designed to explicitly drive the frequency of specific, favorable phenotypes by targeting particular genotypes (U.S. Pat. No. 6,399,855). Fidelity of these associations may be monitored continuously to ensure maintained predictive ability and, thus, informed breeding decisions (US Patent Application 2005/0015827). In this case, costly, time-intensive phenotyping assays required for determining if a plant or plants contains a genomic region associated with a “dicamba tolerance” or “Dicamba intolerance” phenotype can be supplanted by genotypic assays that provide for identification of a plant or plants that contain the desired genomic region that confers dicamba tolerance.

A Genomic Region Associated with a Dicamba Tolerance Phenotype

Provided herewith is a soybean genomic region that is shown herein to be associated with a desirable dicamba tolerance phenotype when present in certain allelic forms and when combined with certain transgenic loci that confer dicamba tolerance.

A soybean genomic region provided that can be associated with a desirable dicamba tolerance phenotype when present in certain allelic forms is located on the telomere proximal end of the short arm of soybean linkage group L (chromosome 19). A series of markers useful in practicing the methods of this invention are provided herewith in Table 1. Additional markers useful in the practice of the invention are provided herewith in Table 2 of the Specification, which is incorporated herewith by reference in its entirety. Table 2 provides the Table 1 markers, additional nucleic acid markers or loci that have been disclosed in various databases, the relative positions of the markers on a physical map of linkage group L (soybean chromosome 19), and sources for the markers.

TABLE 1 Markers spanning a genomic region associated with a desirable dicamba tolerance phenotype Allelic form(s) Marker or SEQ ID Map Associated with Locus Name NO: Position ¹ Dicamba Tolerance ² asmbl_11856 1 16506 TC122822 2 32108 BI967232 3 66686 M0205928 4 92526 M0101742³ 5 112836 TT⁶ M0129138 6 114013 BU551345 7 116147 M0114388 8 380897 BU551363 9 422447 M0205350⁴ 10 423935 TT⁷ M0102027⁵ 11 466558 CC⁸ BU765955 12 474316 M0093116 13 805580 M0129925 14 831128 M0205537 15 890254 M0202715 16 921431 M0206286 17 1209977 M0206054 18 1465354 M0205375 19 2009800 NGMAX008197032⁹ 52 314997 AA¹⁰ ¹ The relative positions of the approximate middle position of the listed markers or loci based on nucleotide positions on a physical map of soybean linkage group L (chromosome 19) of Table 2 are provided where nucleotide position 0 (zero) is telomere proximal and nucleotide position 2009800 is centromere proximal. Polymorphic nucleotide bases are designated in the sequence listing provided herewith according to the WIPO Standard ST.25 (1998), Table 1, as follows: r = g or a (purine); y = t/u or c (pyrimidine); m = a or c; (amino); k = g or t/u (keto); s = g or c (strong interactions 3 H-bonds); w = a or t/u (weak interactions 2H-bonds); b = g or c or t/u (not a); d = a or g or t/u (not c); h = a or c or t/u (not g); v = a or g or c (not t, not u); and n = a or g or c or t/u (unknown, or other; any.) ² Both the maternal and paternal alleles of the single nucleotide polymorphisms that can be associated with a dicamba tolerance phenotype are shown. ³The identified polymorphic allele of marker M0101742 is located at nucleotide 1206 of SEQ ID NO: 5. ⁴The identified polymorphic allele of marker M0205350 is located at nucleotide 148 of SEQ ID NO: 10. ⁵The identified polymorphic allele of marker M0102027is located at nucleotide 349 of SEQ ID NO: 11. ⁶The identified polymorphic allele of marker M0101742 “TT” can be associated with a dicamba tolerance phenotype when the identified polymorphic alleles of the other markers are: “TT” for M0205350 and, in certain embodiments, “CC” for M0102027. ⁷The identified polymorphic allele of marker M020350 “TT” can be associated with a dicamba tolerance phenotype when the identified polymorphic alleles of the other markers are: “TT” for M0101742 and, in certain embodiments, “CC” for M0102027. ⁸In certain embodiments, the identified polymorphic allele “CC” for marker M0102027 can be associated with a dicamba tolerance phenotype when the identified polymorphic alleles of the other markers are: “TT” for M0101742 and “TT” for M020350. ⁹The identified polymorphic allele of marker NGMAX008197032 is located at nucleotide 201 of SEQ ID NO: 52. ¹⁰In certain embodiments, the identified polymorphic allele of marker NGMAX008197032 “AA” can be associated with a dicamba tolerance phenotype when the identified polymorphic alleles of the other markers are: “TT” for M0205350 and, in certain embodiments, “CC” for M0102027, and “TT” for M0101742.

Also provided herein are sub-regions of the linkage group L region that is flanked by loci M0205928 (SEQ ID NO: 4) and BU765995 (SEQ ID NO: 12) that are associated with a dicamba tolerance phenotype. A first sub-region of the linkage group L region associated with a dicamba tolerance phenotype is flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6). These loci flank a first sub-region that spans telomere proximal nucleotide 92334 to centromere proximal nucleotide 113494 in the physical map of linkage group L provided in Table 2 of the specification. Polymorphisms located in this first sub-region that are associated with a dicamba tolerance phenotype can be detected with markers that include, but are not limited to, M0101742 (SEQ ID NO: 5). A second sub-region of the linkage group L region associated with a dicamba tolerance phenotype is flanked by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12). These loci flank the second sub-region that spans telomere proximal nucleotide 422447 to centromere proximal nucleotide 474316 in the physical map of linkage group L provided in Table 2 of the specification. Polymorphisms located in this second sub-region that are associated with a dicamba tolerance phenotype can be detected with markers that include, but are not limited to, M0205350 (SEQ ID NO: 10) or M0102027 (SEQ ID NO: 11). A third sub-region of the linkage group L region associated with a dicamba tolerance phenotype is flanked by loci BU55345 (SEQ ID NO:7) and M0114388 (SEQ ID NO:8). These loci flank the second sub-region that spans telomere proximal nucleotide 115,956 to centromere proximal nucleotide 380,486 in the physical map of linkage group L provided in Table 2 of the specification. Polymorphisms located in this third sub-region that are associated with a dicamba tolerance phenotype can be detected with markers that include, but are not limited to, NGMAX008197032 (SEQ ID NO:52). In certain embodiments of invention, a polymorphism associated with a dicamba tolerant phenotype is detected in only one of these sub-regions. In other embodiments of the invention, at least one polymorphism associated with a dicamba tolerant phenotype is detected in any two of these sub-regions. Thus, a marker including, but not limited to, M0101742 (SEQ ID NO: 5) can be used either independently of, or in combination with, one or more markers selected from the group consisting of M0205350 (SEQ ID NO: 10) and/or M0102027 (SEQ ID NO: 11) to detect polymorphisms associated with a dicamba tolerance phenotype. In certain embodiments, a marker including, but not limited to, M0101742 (SEQ ID NO: 5) can be used either independently of, or in combination with, marker NGMAX008197032 (SEQ ID NO:52) to detect polymorphisms associated with a dicamba tolerance phenotype. In certain embodiments, a marker including, but not limited to, marker NGMAX008197032 (SEQ ID NO:52) can be used either independently of, or in combination with, one or more markers selected from the group consisting of M0205350 (SEQ ID NO: 10) and/or M0102027 (SEQ ID NO: 11) to detect polymorphisms associated with a dicamba tolerance phenotype. In certain embodiments, a polymorphism in the first sub-region is detected with marker M0101742 (SEQ ID NO: 5) and a polymorphism in the second sub-region is detected with markers M0205350 (SEQ ID NO: 10) and/or M0102027 (SEQ ID NO: 11). In certain embodiments, a polymorphism in the first sub-region is detected with marker M0101742 (SEQ ID NO: 5) and a polymorphism in the third sub-region is detected with marker NGMAX008197032 (SEQ ID NO: 52). In certain embodiments, the alleles of these markers associated with dicamba tolerance are a TT allele M0101742 (SEQ ID NO: 5), a TT allele of M0205350 (SEQ ID NO: 10), and, in certain embodiments, a CC allele of M0102027 (SEQ ID NO: 11), and an AA allele of NGMAX008197032 (SEQ ID NO:52).

Additional genetic markers can be used either in conjunction with the markers provided in Table 1 and/or Table 2 or independently of the markers provided in Table 1 and/or Table 2 to practice the methods of the instant invention. Publicly available marker databases from which useful markers can be obtained include, but are not limited to, the soybase.org website on the internet (World Wide Web) that is administered by the United States Agricultural Research Service, the United States Department of Agriculture, and Iowa State University. Additional soybean markers that can be used and that have been described in the literature include, but are not limited to, Hyten et al., BMC Genomics. 11:38, 2010; Choi et al., Genetics. 176(1):685-96, 2007; Yoon et al., Theor Appl Genet. 2007 March; 114(5):885-99; and Hyten et al. Crop Sci. 2010 50: 960-968. Given the provision herein of a genomic region on linkage group L (chromosome 19) delimited or flanked by the telomere proximal locus M0205928 (SEQ ID NO: 4) of Table 2 and the centromere proximal locus BU765955 (SEQ ID NO: 12) of Table 2 as well as an assortment of soybean germplasms exhibiting either a “dicamba intolerant” or “dicamba tolerant” phenotype, additional markers located either within or near this genomic region that are associated with these phenotypes can be obtained by merely typing the new markers in the various germplasms provided herewith. The genomic region on linkage group L (chromosome 19) delimited or flanked by the telomere proximal locus M0205928 (SEQ ID NO: 5) of Table 2 and the centromere proximal locus BU765955 (SEQ ID NO: 12) of Table 2 can also be mapped relative to markers provided in any publicly available or other soybean physical or genetic map to place this genetic locus on that map.

Identification of Plants Exhibiting the “Dicamba Intolerance” or “Dicamba Tolerance” Phenotype

To observe the presence or absence of the “dicamba intolerance” or dicamba tolerance phenotypes, transgenic soybean plants comprising a transgene that confers resistance to dicamba are typically exposed in early to mid-vegetative growth stages to one or more high doses of dicamba. Typical doses of dicamba that can elicit a dicamba intolerance phenotype can range from about a 2-fold label application rate of a commercially available dicamba formulation to about a 3-fold label application rate of a commercially available dicamba formulation. In terms of acid equivalents of dicamba acid applied, typical doses of dicamba that can elicit a dicamba intolerance phenotype can range from an application rate of about 1.0 pounds of acid equivalent per acre of dicamba acid to about 1.5 pounds of acid equivalent per acre of dicamba acid when the indicated amounts of dicamba acid are provided in either a commercially available dicamba formulation or when the indicated amounts of dicamba acid is provided in a similar formulation suitable for application to dicamba-tolerant crops. Commercially available dicamba formulations that can be used include, but are not limited to, Clarity® (BASF, N.C., USA); Banvel®, Banvel M®, Banvel Banvel SGF®, or Vanquish® (Syngenta, Wilmington, Del., USA); or Rifle® (Loveland Products, Inc., Loveland, Colo., USA). In certain embodiments, the commercially available dicamba formulation used is Clarity®. In certain embodiments, doses of dicamba that can elicit a dicamba intolerance phenotype can range from about a 2 fold application rate of about 0.25 gallons per acre Clarity® to about a three fold application rate of about 0.375 gallons per acre per acre Clarity®.

The dicamba intolerance phenotype can be observed approximately a week after herbicide application in certain soybean varieties comprising the transgene that confers resistance to dicamba. Dicamba is typically applied during pre and post-emergent vegetative growth stages. In certain embodiments of these methods, dicamba can be applied in weekly intervals (i.e. once a week) for any of 2, 3, 4 or more successive weeks to score for the presence of the dicamba intolerance phenotype. In certain embodiments, soybean plants at about the V3 vegetative development stage are exposed to an initial dicamba spray followed by a subsequent spray at V6/R1. Genotypes provided herein are especially useful for providing dicamba tolerance to plants sprayed at the V6 stage. As discussed herein, the vegetative stages of soybean are as follows: VE (emergence), VC (cotyledon stage), V1 (first trifoliate leaf), V2 (second trifoliate leaf), V3 (third trifoliate leaf), V(n) (nth trifoliate leaf), and V6 (flowering will soon start). As discussed herein, the reproductive stages of soybean are as follows: R1 (beginning bloom), R2 (full bloom), R3 (beginning pod), R4 (full pod), R5 (beginning seed), R6 (full seed), R7 (beginning maturity) and R8 (full maturity). A description of the soybean vegetative and reproductive stages can be found on the world wide web (internet) at ag.ndsu.edu/pubs/plantsci/rowcrops/a1174/a1174 w.htm (North Dakota State University publication A-1174, June 1999, Reviewed and Reprinted August 2004).

A rating scale that evaluates the degree of dicamba intolerance can also be employed to identify “dicamba intolerant” and “dicamba tolerant” plants. An exemplary and non-limiting scale for evaluating the Dicamba intolerance phenotype is as follows, where a low number corresponds to a “dicamba tolerance” phenotype and the a high number correlates to a “dicamba intolerance” phenotype:

A rating of 1: Less than 10% of plants show malformation A rating of 2: 10-50% of plants show malformation A rating of 3: Greater than 50% of plants show malformation

Identification of Plants Exhibiting Reproductive Tolerance to Glyphosate Phenotype

To observe the presence or absence of reproductive tolerance to glyphosate phenotypes, transgenic soybean plants comprising a transgene that confers glyphosate resistance are typically exposed in mid- to late-vegetative growth stages to one or more high doses of glyphosate. Doses of glyphosate that can elicit a reproductive sensitivity phenotype are usually at least about twice the typical application rates of commercial glyphosate formulations that are used to provide weed control in transgenic, glyphosate resistant soybean plants. In terms of acid equivalents of glyphosate acid applied, typical doses of glyphosate that can elicit a reproductive sensitivity phenotype can range from an application rate of about 1.0 pounds of acid equivalent per acre (about 1.12 kilograms per hectare) of glyphosate acid to about 2.25 pounds of acid equivalent per acre (i.e. about 2.52 kilograms per hectare) of glyphosate acid when the indicated amounts of glyphosate acid are provided in either a commercially available glyphosate formulation or when the indicated amounts of glyphosate acid is provided in a similar formulation suitable for application to glyphosate-tolerant crops. Commercially available glyphosate formulations that can be used include, but are not limited to, Roundup Original MAX®, Roundup PowerMAX®, Roundup UltraMax®, or RoundUp WeatherMAX® (Monsanto Co., St. Louis, Mo., USA); Touchdown IQ® or Touchdown Total® (Syngenta, Wilmington, Del., USA); Glyphomax®, Glyphomax Plus®, or Glyphomax XRT® (Dow Agrosciences LLC, Indianapolis, Ind., USA). In certain embodiments, the commercially available glyphosate formulation used is RoundUp WeatherMAX®. In certain embodiments, doses of glyphosate that can elicit a reproductive sensitivity phenotype can range from about a 2 fold application rate of about 42.6 ounces per acre RoundUp WeatherMax® (1.68 kilograms per hectare) to about a three fold application rate of about 63.9 ounces per acre RoundUp WeatherMax® (i.e. about 2.52 kilograms per hectare).

The reproductive sensitivity phenotype can be observed at an appropriate stage of reproductive development after herbicide application in certain soybean varieties comprising the transgene that confers glyphosate resistance. Glyphosate is typically applied during vegetative growth stages, where applications in later vegetative growth stages can typically elicit reproductive sensitivity at lower application rates. In certain embodiments of these methods, glyphosate can be applied in weekly intervals (i.e. once a week) for any of 2, 3, 4 or more successive weeks to score for the presence of the reproductive sensitivity phenotype. In certain embodiments, soybean plants at about the V3 vegetative development stage are exposed to an initial glyphosate spray followed by a subsequent spray at the V6 vegetative stage. In certain embodiments, soybean plants at about the V6 vegetative development stage are exposed to a glyphosate spray. As discussed herein, the vegetative stages of soybean are as follows: VE (emergence), VC (cotyledon stage), V1 (first trifoliolate leaf), V2 (second trifoliolate leaf), V3 (third trifoliolate leaf), V(n) (nth trifoliolate leaf), and V6 (flowering will soon start). As discussed herein, the reproductive stages of soybean are as follows R1 (beginning bloom, first flower); R2 (full bloom, flower in top 2 nodes); R3 (beginning pod, 3/16″ pod in top 4 nodes); R4 (full pod, ¾″ pod in top 4 nodes); R5 (⅛″ seed in top 4 nodes); R6 (full size seed in top 4 nodes); R7 (beginning maturity, one mature pod); and, R8 (full maturity, 95% of pods on the plant are mature). A description of the soybean vegetative and reproductive stages can be found on the world wide web (internet) at ag.ndsu.edu/pubs/plantsci/rowcrops/a1174/a1174w.htm (North Dakota State University publication A-1174, June 1999, Reviewed and Reprinted August 2004). Expression of the reproductive sensitivity trait can also be influenced by temperature, where the trait in varieties that display the reproductive sensitivity phenotype is more pronounced following treatment at temperatures of about 32 degrees Celsius or more.

A rating scale that evaluates the degree of reproductive sensitivity can also be employed to identify “tolerant” and “sensitive” plants. An exemplary and non limiting scale for evaluating the reproductive sensitivity phenotype is as follows, where the low numbers correspond to a “glyphosate reproductive tolerance” phenotype and the high numbers correlate to a “glyphosate reproductive sensitivity” phenotype where sterility is monitored as follows:

-   -   A rating scale of 1: Less than 10% of plants show sterility         (glyphosate reproductive tolerance)     -   A rating scale of 2: Less than 10-50% of plants show sterility     -   A rating scale 3: Greater than 50% of plants show sterility         (glyphosate reproductive sensitivity)         Soybean plant sterility can be measured by a variety of methods         that include, but are not limited to, determining pollen counts,         seed yield per plant, seed yield per pod, and the like. Controls         used to determine glyphosate reproductive sensitivity or         tolerance of a given transgenic soybean test plant comprising a         transgenic insertion event that confers glyphosate resistance in         a certain genetic background (i.e. genotype) in comparison tests         include, but are not limited to, (a) co-cultivated soybean         plants comprising the same transgenic insertion event in a         genetic background that provides for glyphosate reproductive         sensitivity; and/or (b) co-cultivated soybean plants comprising         the same transgenic insertion event in a genetic background that         provides for glyphosate reproductive tolerance, where the test         and control plants are sprayed with glyphosate. Additional         controls used to determine glyphosate reproductive sensitivity         or tolerance can also include, but are not limited to,         co-cultivated soybean plants of the same genotypes (i.e. soybean         plants that are isogenic with respect to both the transgenic         insertion event and genetic background as either the test or         control soybean lines) that are not sprayed with glyphosate.         Introgression of a Genomic Region Associated with a Dicamba         Tolerance Phenotype

Also provided herewith is unique soybean germplasm comprising an introgressed genomic region that is associated with a dicamba tolerance phenotype and methods of obtaining the same. Marker-assisted introgression involves the transfer of a chromosomal region, defined by one or more markers, from one germplasm to a second germplasm. Offspring of a cross that contain the introgressed genomic region can be identified by the combination of markers characteristic of the desired introgressed genomic region from a first germplasm (i.e. such as a dicamba tolerance germplasm) and both linked and unlinked markers characteristic of the desired genetic background of a second germplasm (i.e. a dicamba intolerance germplasm). In addition to the markers provided herewith that identify alleles of genomic region that is associated with a dicamba tolerance phenotype, flanking markers that fall on both the telomere proximal end of the genomic region on linkage group L (chromosome 19) and the centromere proximal end of the linkage group L (chromosome 19) genomic region are also provided in Tables 1 and 2. Table 2 is provided at the end of the specification immediately before the claims. Such flanking markers are useful in a variety of breeding efforts that include, but are not limited to, introgression of the genomic region associated with a dicamba tolerance phenotype into a genetic background comprising markers associated with germplasm that ordinarily contains the allelic forms of the genomic region that is associated with a “Dicamba intolerance” phenotype. Telomere proximal flanking markers that can be used in these methods include, but are not limited to, asmbl_11856 (SEQ ID NO: 1), TC122822 (SEQ ID NO: 2), BI967232 (SEQ ID NO: 3), and/or polymorphisms in any of the loci listed in Table 2 of the Specification located between starting base 16426 (the telomere proximal base) of locus asmbl_11856 and starting base 92334 of locus M0205928 (SEQ ID NO: 4). Centromere proximal flanking markers that can be used in these methods include, but are not limited to, M0205537 (SEQ ID NO: 15), M0202715 (SEQ ID NO: 16), M0206286 (SEQ ID NO: 17), M0206054 (SEQ ID NO: 18) and M0205375 (SEQ ID NO: 19) and/or polymorphisms in any of the other loci listed in Table 2 that are centromere proximal to BU765955 (SEQ ID NO: 12). Soybean plants wherein the two subregions that are respectively flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6 and by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12) are selectively introgressed can be obtained by using the BU551345 (SEQ ID NO: 7), SATT723, and/or M0114388 (SEQ ID NO: 8) markers, or by using any of the markers located between these two subregions that are provided in Table 2. Any of the aforementioned polymorphisms can be identified by sequencing loci from dicamba intolerant and dicamba tolerance germplasms. Additional markers located on linkage group L (chromosome 19) and other chromosomes are disclosed in US Patent Application Publication 20090208964. Publicly available marker databases from which additional useful markers located on linkage group L (chromosome 19) and other chromosomes can be obtained include, but are not limited to, the soybase.org website on the internet that is administered by the United States Agricultural Research Service, the United States Department of Agriculture, and Iowa State University. Soybean plants or germplasm comprising an introgressed genomic region that is associated with a dicamba tolerance phenotype wherein at least 10%, 25%, 50%, 75%, 90%, or 99% of the remain genomic sequences carry markers characteristic of soybean plants or germplasm that are otherwise or ordinarily comprise a genomic region associated with the Dicamba intolerance phenotype are thus provided.

Soybean Plants Comprising Genomic Region Associated with the Dicamba Intolerance and Dicamba Tolerance Phenotypes and Transgenes that Confer Resistance to Dicamba

A non-limiting and exemplary list of soybean plants that comprise genomic regions associated with either a dicamba-intolerance or a dicamba tolerance phenotype are provided herewith in Table 3.

TABLE 3 Soybean varieties comprising a genomic region associated with a dicamba tolerance or dicamba intolerant phenotype. ATCC Variety Depository Date of Branded Dicamba U.S. Pat. Name in Accession Patent Name ¹ Phenotype No. Patent Number ² Issue AG3102 Intolerant 7,964,777 7629164 PTA-10825 Jun. 21, 2011 AG3603 Intolerant 7,592,516 D4328762 PTA-9797 Sep. 22, 2009 AG4903 AG4907 Intolerant 7,687,685 D5703684 PTA-10153 Mar. 30, 2010 AG0803 Tolerant 7,498,489 4498438 PTA-9064 AG3102 Tolerant 7,964,777 7629164 PTA-10825 Jun. 21, 2011 AG3603 Tolerant 7,592,516 D4328762 PTA-9797 Sep. 22, 2009 BBL3606N0R BL3307M2-D0RL AG4903 AG4907 Tolerant 7,687,685 D5703684 PTA-10153 Mar. 30, 2010 260744-14 AFL0506C0R Tolerant 7,723,583 D5864369 PTA-10719 May 25, 2010 AG0808 Tolerant 7,732,672 D5142326 PTA-10168 Jun. 8, 2010 263619-24 4065735-51 5463213-25 AG1002 Tolerant 7,294,770 5826175 PTA-8148 Nov. 13, 2007 AG1403 Tolerant 7,557,273 6943322 PTA-9554 Jul. 7, 2009 AG1406 Tolerant 7,732,673 D5232589 PTA-10268 Jun. 8, 2010 CSR1920 Tolerant 7,728,199 7821295 PTA-10519 Jun. 1, 2010 15733-79-59 5081541-27 5464705-06 AG2110 Tolerant 7,678,965 D5624834 PTA-10134 Mar. 16, 2010 AG2606 Tolerant 7,622,644 D4201139 PTA-9749 Nov. 24, 2009 AG2909 Tolerant 7,999,153 D5502014 PTA-11081 Aug. 16, 2011 AG2921V Tolerant 7,390,940 4858197 PTA-9072 Jun. 24, 2008 AG3021V Tolerant 7,572,958 D4361423 PTA-9801 Aug. 11, 2009 BOX2906H0R DFN3306B0R Tolerant 7,626,089 D4311702 PTA-9781 Dec. 1, 2009 CSRS4782N Tolerant 7,700,847 D5898941 PTA-10598 Apr. 20, 2010 GL4807A2-D0RN Tolerant 7,868,230 D5523145 PTA-11362 Jan. 11, 2011 ¹ Branded names of Asgrow ® (designated “AG”) and DEKALB ® soybean varieties from Monsanto Co. 800 N. Lindbergh Blvd., St. Louis, MO, USA. ² Deposit numbers of seed available through the American Type Culture Collection (ATCC), 10801 University Blvd., Manassas, Va., USA, 20110-2209. ³ Dicamba phenotype is the phenotype observed in the indicated germplasm containing a transgene that confers resistance to dicamba when exposed to dicamba.

Also provided herewith are additional soybean plants that comprising a genomic region associated with a dicamba intolerant or dicamba tolerance phenotype that are identified by use of the markers provided in Table 1 and/or Table 2 and/or methods provided herein. Any of the soybean plants identified in Table 3 or other soybean plants that are otherwise identified using the markers or methods provided herein can be used in methods that include, but are not limited to, methods of obtaining soybean plants with an introgressed dicamba tolerance locus, obtaining a soybean plant that exhibits a dicamba tolerance phenotype, or obtaining a soybean plant comprising in its genome a genetic region associated with a dicamba tolerance phenotype.

In certain embodiments, the soybean plants provided herein or used in the methods provided herein can comprise a transgene that confers resistance to dicamba. In certain embodiments, the dicamba tolerant soybean plants can comprise a transgene encoding a dicamba-degrading dicamba monoxygenase (DMO) enzyme that catalyzes the conversion of herbicidal dicamba (3, 6-dichloro-o-anisic acid) to a non-toxic 3, 6-dichlorosalicylic acid. In certain embodiments, the dicamba-degrading dicamba monoxygenase (DMOw) comprise a DMO enzyme disclosed in U.S. Pat. Nos. 7,022,896, 7,105,724, and 7,812,224, each incorporated herein by reference in their entireties. Exemplary and non-limiting DMOw dicamba monooxygenase encoding nucleic acid and protein sequences are provided herewith as SEQ ID NO: 20 and SEQ ID NO: 21. In certain embodiments, the dicamba tolerant soybean plants can comprise a dicamba monoxygenase variant which exhibits improved catalytic parameters such as increased turnover number and/or a lower km for the substrate, improved catalysis at lower pH values, and/or improved catalysis at higher temperatures relative to an unaltered dicamba monooxygenase. In certain embodiments, the dicamba monoxygenase variant comprises a DMOc variant enzyme disclosed in U.S. Pat. No. 7,884,262, incorporated herein by reference in its entirety. Exemplary and non-limiting DMOc dicamba monooxygenase variant encoding nucleic acid and protein sequences are provided herewith as SEQ ID NO: 22 and SEQ ID NO: 23. In certain embodiments, a dicamba monooxygenase is operably linked to a chloroplast transit peptide (CTP). Operable linkage of certain CTPs to DMO is disclosed in U.S. Pat. No. 8,084,666, which is incorporated herein by reference in its entirety. In certain embodiments, it is contemplated that the soybean plants used herein can comprise one or more specific genomic insertion(s) of a dicamba tolerant transgene including, but not limited to, as those found in MON87708 soybean (deposited under ATCC accession number PTA-9670 and described in US Patent Application Publication Number 20110067134).

In certain embodiments, the soybean plants provided herein or used in the methods provided herein can comprise a transgene that confers tolerance to glyphosate. Transgenes that can confer tolerance to glyphosate include, but are not limited to, transgenes that encode glyphosate tolerant Class I EPSPS (5-enolpyruvylshikimate-3-phosphate synthases) enzymes or glyphosate tolerant Class II EPSPS (5-enolpyruvylshikimate-3-phosphate synthases) enzymes. Useful glyphosate tolerant EPSPS enzymes provided herein are disclosed in U.S. Pat. Nos. 6,803,501, RE39,247, 6,225,114, 5,188,642, and 4,971,908. In certain embodiments, the glyphosate tolerant soybean plants can comprise a transgene encoding a glyphosate oxidoreductase or other enzyme which degrades glyphosate. Glyphosate oxidoreductase enzymes had been described in U.S. Pat. No. 5,776,760 and US Reissue patent RE38,825. In certain embodiments the soybean plant can comprise a transgene encoding a glyphosate N-acetyltransferase gene that confers tolerance to glyphosate. In certain embodiments, the soybean plant can comprise a glyphosate n-acetyltransferase encoding transgene such as those described in U.S. Pat. No. 7,666,644. In still other embodiments, soybean plants comprising combinations of transgenes that confer glyphosate tolerance are provided. Soybean plants comprising both a glyphosate resistant EPSPS and a glyphosate N-acetyltransferase are also provided herewith. In certain embodiments, it is contemplated that the soybean plants used herein can comprise one or more specific genomic insertion(s) of a glyphosate tolerant transgene including, but not limited to, as those found in: i) MON89788 soybean (deposited under ATCC accession number PTA-6708 and described in US Patent Application Publication Number 20100099859), ii) GTS 40-3-2 soybean (Padgette et al., Crop Sci. 35: 1451-1461, 1995), iii) event 3560.4.3.5 soybean (seed deposited under ATCC accession number PTA-8287 and described in US Patent Publication 20090036308), or any combination of i (MON89788 soybean), ii (GTS 40-3-2 soybean), and iii (event 3560.4.3.5 soybean).

In certain embodiments, the gene that confers resistance to dicamba is a gene encoding a Dicamba monooxygenase (DMO). The DMO gene is a microbial gene that has been transformed into soybean and cotton to confer tolerance to the dicamba herbicide. The DMO protein expressed in the plants transformed with the DMO gene actively metabolizes dicamba to 3,6-dichloro salicylic acid (DCSA), which lacks herbicidal activity. In certain embodiments, Dicamba resistant (DR) soybeans can be crossed with “RoundUp Ready 2 Yield™” (RR2Y) soybeans to generate a stack (RR2YxDR) which can confer resistance to both dicamba and glyphosate. It has been observed in certain germplasms that a herbicide traits (i.e. transgene conferred glyphosate and dicamba resistance)×germplasm interaction can result in increased sensitivity to dicamba (i.e. “dicamba intolerance”) that may be commercially undesirable. In certain embodiments, favorable haplotypes are provided herein which are associated with robust tolerance to dicamba and glyphosate, and which are useful for selection of RR2YxDR soybeans that do not exhibit dicamba intolerance.

In certain embodiments, it is contemplated that genotypic assays that provide for non-destructive identification of the plant or plants can be performed either in seed, the emergence stage, the “VC” stage (i.e. cotyledons unfolded), the V1 stage (appearance of first node and unifoliate leaves), the V2 stage (appearance of the first trifoliate leaf), and thereafter. In certain embodiments, non-destructive genotypic assays are performed in seed using apparati and associated methods as described in U.S. Pat. Nos. 6,959,617; 7,134,351; 7,454,989; 7,502,113; 7,591,101; 7,611,842; and 7,685,768, which are incorporated herein by reference in their entireties. In certain embodiments, non-destructive genotypic assays are performed in seed using apparati and associated methods as described in US Patent Application Publications 20100086963, 20090215060, and 20090025288, which are incorporated herein by reference in their entireties. Published U.S. Patent Applications US 2006/0042527, US 2006/0046244, US 2006/0046264, US 2006/0048247, US 2006/0048248, US 2007/0204366, and US 2007/0207485, which are incorporated herein by reference in their entirety, also disclose apparatus and systems for the automated sampling of seeds as well as methods of sampling, testing and bulking seeds. Thus, in a certain embodiments, any of the methods provided herein can comprise screening for markers in individual seeds of a population wherein only seed with at least one genotype of interest is advanced.

Soybean Plants Comprising a Genomic Region Associated with Stacked Transgenic Trait Improvement and Transgenes that Confer Resistance to Other Herbicides and/or Insects

In certain embodiments, soybean plants comprising a genomic region associated with stacked transgenic trait improvement (or the dicamba tolerance phenotype) and at least one additional herbicide resistance transgene selected from the group consisting of a dicamba resistance conferring transgene, a 2,4-D resistance conferring transgene, and a glufosinate resistance conferring transgene and/or at least one transgene encoding a product that confers insect resistance selected from the group consisting of a dsRNA that inhibits a target gene of an insect pest, a patatin, a Bacillus thuringiensis insecticidal protein, a Xenorhabdus insecticidal protein, a Photorhabdus insecticidal protein, a Bacillus laterosporous insecticidal protein, a Bacillus sphaericus insecticidal protein, and a lignin are provided herein. Such transgenic trait improvements that can occur in plants comprising the genomic regions provided herein can be ascertained by comparing transgenic trait performance in varieties containing the genomic regions to the transgenic trait performance in other varieties lacking the genomic region. Such transgenic herbicide resistance trait improvements that can occur in plants comprising the genomic regions provided herein can include, but are not limited to, decreased phytotoxicity upon herbicide exposure in varieties containing the genomic regions conferring the improved transgenic trait performance and the corresponding herbicide resistance conferring transgene in comparison to other varieties lacking the genomic region and the corresponding herbicide resistance conferring transgene upon herbicide exposure. Various dsRNAs that inhibit a target gene of an insect pest are described in US Patent Application Publication Number 20120137387, which is specifically incorporated herein by reference in its entirety. A Bacillus thuringiensis insecticidal protein can be any of a number of insecticidal proteins including but not limited to a Cry1, a Cry3, a TIC851, a CryET70, a Cry22, a TIC901, a TIC1201, a TIC407, a TIC417, a binary insecticidal protein CryET33 and CryET34, a binary insecticidal protein CryET80 and CryET76, a binary insecticidal protein TIC100 and TIC101, a binary insecticidal protein PS149B1, a VIP insecticidal protein, a TIC900 or related protein, or combinations of the insecticidal proteins ET29 or ET37 with insecticidal proteins TIC810 or TIC812, and insecticidal chimeras of any of the preceding insecticidal proteins. A Bacillus thuringiensis insecticidal protein can be any of a number of insecticidal proteins including but not limited to a Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ad, Cry1Ae, Cry1Ba, Cry1Bb, Cry1Ca, Cry1Cb, Cry1Da, Cry1Db, Cry1Ea, Cry1Eb, Cry1Fa, Cry1Fb, Cry1Ga, Cry1Ha, Cry2Aa, Cry2Ab, Cry1Ja, Cry1Ka, Cry11Aa, Cry11Ab, Cry12Aa, Cry3Ba, Cry3Bb, Cry3C, Cry4a, Cry4Ba, Cry5a, Cry5Ab, Cry6Aa, Cry6Ba, Cry7Aa, Cry7Ab, Cry8Aa, Cry8Ba, Cry8Ca, Cry9Aa, Cry9Ba, Cry9Ca, Cry10Aa, Cry11Aa, Cry12Aa, Cry13Aa, Cry14Aa, Cry15Aa, Cyt1Aa, and Cyt2Aa protein or an insecticidal chimeras thereof. Insecticidal chimeras of certain Bacillus thuringiensis insecticidal proteins include, but are not limited to, Cry1A/F and other chimeras disclosed in US Patent Application Publication No. 20090143298. Such transgenic insect resistance trait improvements that can occur in plants comprising the genomic regions provided herein can include, but are not limited to, decreased insect-mediated plant damage, or increased insect death, inhibition, stunting, or cessation of insect feeding in varieties containing the genomic regions that confer the transgenic trait performance in comparison to other varieties lacking the genomic region.

Molecular Assisted Breeding Techniques

Genetic markers that can be used in the practice of the instant invention include, but are not limited to, are Restriction Fragment Length Polymorphisms (RFLP), Amplified Fragment Length Polymorphisms (AFLP), Simple Sequence Repeats (SSR), Single Nucleotide Polymorphisms (SNP), Insertion or Deletion Polymorphisms (Indels), Variable Number Tandem Repeats (VNTR), and Random Amplified Polymorphic DNA (RAPD), and others known to those skilled in the art. Marker discovery and development in crops provides the initial framework for applications to marker-assisted breeding activities (US Patent Applications 2005/0204780, 2005/0216545, 2005/0218305, and 2006/00504538). The resulting “genetic map” is the representation of the relative position of characterized loci (DNA markers or any other locus for which alleles can be identified) along the chromosomes. The measure of distance on this map is relative to the frequency of crossover events between sister chromatids at meiosis.

As a set, polymorphic markers serve as a useful tool for fingerprinting plants to inform the degree of identity of lines or varieties (U.S. Pat. No. 6,207,367). These markers form the basis for determining associations with phenotype and can be used to drive genetic gain. The implementation of marker-assisted selection is dependent on the ability to detect underlying genetic differences between individuals.

Certain genetic markers for use in the present invention include “dominant” or “codominant” markers. “Codominant markers” reveal the presence of two or more alleles (two per diploid individual). “Dominant markers” reveal the presence of only a single allele. The presence of the dominant marker phenotype (e.g., a band of DNA) is an indication that one allele is present in either the homozygous or heterozygous condition. The absence of the dominant marker phenotype (e.g., absence of a DNA band) is merely evidence that “some other” undefined allele is present. In the case of populations where individuals are predominantly homozygous and loci are predominantly dimorphic, dominant and codominant markers can be equally valuable. As populations become more heterozygous and multiallelic, codominant markers often become more informative of the genotype than dominant markers.

In another embodiment, markers that include, but are not limited to, single sequence repeat markers (SSR), AFLP markers, RFLP markers, RAPD markers, phenotypic markers, isozyme markers, single nucleotide polymorphisms (SNPs), insertions or deletions (Indels), single feature polymorphisms (SFPs, for example, as described in Borevitz et al. 2003 Gen. Res. 13:513-523), microarray transcription profiles, DNA-derived sequences, and RNA-derived sequences that are genetically linked to or correlated with dicamba tolerance loci, regions flanking dicamba tolerance loci, regions linked to dicamba tolerance loci, and/or regions that are unlinked to dicamba tolerance loci can be used in certain embodiments of the instant invention.

In one embodiment, nucleic acid-based analyses for determining the presence or absence of the genetic polymorphism (i.e. for genotyping) can be used for the selection of seeds in a breeding population. A wide variety of genetic markers for the analysis of genetic polymorphisms are available and known to those of skill in the art. The analysis may be used to select for genes, portions of genes, QTL, alleles, or genomic regions (Genotypes) that comprise or are linked to a genetic marker that is linked to or correlated with dicamba tolerance loci, regions flanking dicamba tolerance loci, regions linked to dicamba tolerance loci, and/or regions that are unlinked to dicamba tolerance loci can be used in certain embodiments of the instant invention.

Nucleic acid analysis methods provided herein include, but are not limited to, PCR-based detection methods (for example, TaqMan™ assays), microarray methods, mass spectrometry-based methods and/or nucleic acid sequencing methods. In one embodiment, the detection of polymorphic sites in a sample of DNA, RNA, or cDNA may be facilitated through the use of nucleic acid amplification methods. Such methods specifically increase the concentration of polynucleotides that span the polymorphic site, or include that site and sequences located either distal or proximal to it. Such amplified molecules can be readily detected by gel electrophoresis, fluorescence detection methods, or other means.

A method of achieving such amplification employs the polymerase chain reaction (PCR) (Mullis et al. 1986 Cold Spring Harbor Symp. Quant. Biol. 51:263-273; European Patent 50,424; European Patent 84,796; European Patent 258,017; European Patent 237,362; European Patent 201,184; U.S. Pat. Nos. 4,683,202; 4,582,788; and 4,683,194), using primer pairs that are capable of hybridizing to the proximal sequences that define a polymorphism in its double-stranded form.

Methods for typing DNA based on mass spectrometry can also be used. Such methods are disclosed in U.S. Pat. Nos. 6,613,509 and 6,503,710, and references found therein. Polymorphisms in DNA sequences can be detected or typed by a variety of effective methods well known in the art including, but not limited to, those disclosed in U.S. Pat. Nos. 5,468,613, 5,217,863; 5,210,015; 5,876,930; 6,030,787; 6,004,744; 6,013,431; 5,595,890; 5,762,876; 5,945,283; 5,468,613; 6,090,558; 5,800,944; 5,616,464; 7,312,039; 7,238,476; 7,297,485; 7,282,355; 7,270,981 and 7,250,252 all of which are incorporated herein by reference in their entireties. However, the compositions and methods of the present invention can be used in conjunction with any polymorphism typing method to type polymorphisms in genomic DNA samples. These genomic DNA samples used include but are not limited to genomic DNA isolated directly from a plant, cloned genomic DNA, or amplified genomic DNA.

For instance, polymorphisms in DNA sequences can be detected by hybridization to allele-specific oligonucleotide (ASO) probes as disclosed in U.S. Pat. Nos. 5,468,613 and 5,217,863. U.S. Pat. No. 5,468,613 discloses allele specific oligonucleotide hybridizations where single or multiple nucleotide variations in nucleic acid sequence can be detected in nucleic acids by a process in which the sequence containing the nucleotide variation is amplified, spotted on a membrane and treated with a labeled sequence-specific oligonucleotide probe.

Target nucleic acid sequence can also be detected by probe ligation methods as disclosed in U.S. Pat. No. 5,800,944 where sequence of interest is amplified and hybridized to probes followed by ligation to detect a labeled part of the probe.

Microarrays can also be used for polymorphism detection, wherein oligonucleotide probe sets are assembled in an overlapping fashion to represent a single sequence such that a difference in the target sequence at one point would result in partial probe hybridization (Borevitz et al., Genome Res. 13:513-523 (2003); Cui et al., Bioinformatics 21:3852-3858 (2005). On any one microarray, it is expected there will be a plurality of target sequences, which may represent genes and/or noncoding regions wherein each target sequence is represented by a series of overlapping oligonucleotides, rather than by a single probe. This platform provides for high throughput screening a plurality of polymorphisms. A single-feature polymorphism (SFP) is a polymorphism detected by a single probe in an oligonucleotide array, wherein a feature is a probe in the array. Typing of target sequences by microarray-based methods is disclosed in U.S. Pat. Nos. 6,799,122; 6,913,879; and 6,996,476.

Target nucleic acid sequence can also be detected by probe linking methods as disclosed in U.S. Pat. No. 5,616,464, employing at least one pair of probes having sequences homologous to adjacent portions of the target nucleic acid sequence and having side chains which non-covalently bind to form a stem upon base pairing of the probes to the target nucleic acid sequence. At least one of the side chains has a photoactivatable group which can form a covalent cross-link with the other side chain member of the stem.

Other methods for detecting SNPs and Indels include single base extension (SBE) methods. Examples of SBE methods include, but are not limited, to those disclosed in U.S. Pat. Nos. 6,004,744; 6,013,431; 5,595,890; 5,762,876; and 5,945,283. SBE methods are based on extension of a nucleotide primer that is adjacent to a polymorphism to incorporate a detectable nucleotide residue upon extension of the primer. In certain embodiments, the SBE method uses three synthetic oligonucleotides. Two of the oligonucleotides serve as PCR primers and are complementary to sequence of the locus of genomic DNA which flanks a region containing the polymorphism to be assayed. Following amplification of the region of the genome containing the polymorphism, the PCR product is mixed with the third oligonucleotide (called an extension primer) which is designed to hybridize to the amplified DNA adjacent to the polymorphism in the presence of DNA polymerase and two differentially labeled dideoxynucleosidetriphosphates. If the polymorphism is present on the template, one of the labeled dideoxynucleosidetriphosphates can be added to the primer in a single base chain extension. The allele present is then inferred by determining which of the two differential labels was added to the extension primer. Homozygous samples will result in only one of the two labeled bases being incorporated and thus only one of the two labels will be detected. Heterozygous samples have both alleles present, and will thus direct incorporation of both labels (into different molecules of the extension primer) and thus both labels will be detected.

In another method for detecting polymorphisms, SNPs and Indels can be detected by methods disclosed in U.S. Pat. Nos. 5,210,015; 5,876,930; and 6,030,787 in which an oligonucleotide probe having a 5′ fluorescent reporter dye and a 3′ quencher dye covalently linked to the 5′ and 3′ ends of the probe. When the probe is intact, the proximity of the reporter dye to the quencher dye results in the suppression of the reporter dye fluorescence, e.g. by Forster-type energy transfer. During PCR forward and reverse primers hybridize to a specific sequence of the target DNA flanking a polymorphism while the hybridization probe hybridizes to polymorphism-containing sequence within the amplified PCR product. In the subsequent PCR cycle DNA polymerase with 5′→3′ exonuclease activity cleaves the probe and separates the reporter dye from the quencher dye resulting in increased fluorescence of the reporter.

In another embodiment, the locus or loci of interest can be directly sequenced using nucleic acid sequencing technologies. Methods for nucleic acid sequencing are known in the art and include technologies provided by 454 Life Sciences (Branford, Conn.), Agencourt Bioscience (Beverly, Mass.), Applied Biosystems (Foster City, Calif.), LI-COR Biosciences (Lincoln, Nebr.), NimbleGen Systems (Madison, Wis.), Illumina (San Diego, Calif.), and VisiGen Biotechnologies (Houston, Tex.). Such nucleic acid sequencing technologies comprise formats such as parallel bead arrays, sequencing by ligation, capillary electrophoresis, electronic microchips, “biochips,” microarrays, parallel microchips, and single-molecule arrays, as reviewed by R.F. Service Science 2006 311:1544-1546.

The markers to be used in the methods of the present invention should preferably be diagnostic of origin in order for inferences to be made about subsequent populations. Experience to date suggests that SNP markers may be ideal for mapping because the likelihood that a particular SNP allele is derived from independent origins in the extant populations of a particular species is very low. As such, SNP markers appear to be useful for tracking and assisting introgression of QTLs, particularly in the case of Genotypes.

EXAMPLES

The following examples are included to demonstrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention.

Example 1

Marker-assisted backcrossing (MABC) is a common breeding methodology to transfer a gene of interest into a desired recurrent parent. MABC was used to transfer both the dicamba resistance (DMO) transgene (U.S. Patent Appl. US20110067134) and the glyphosate resistant RoundUp Ready 2 Yield™ (RR2Y) CP4 genes (U.S. Pat. No. 7,632,985) into several recurrent parents. The process involved making three backcrosses to the recurrent parent and using genome wide markers to target the recovery of 95% or greater of the recurrent parent genome. In this process, markers were used to confirm the presence of the DMO and RR2Y CP4 genes and the absence of the RR1 CP4 gene. The haplotype at the dicamba tolerance locus for each recurrent parent was known.

Observations on MABC lines whose recurrent parents contained different haplotypes were made. For each MABC, there were several variants, with each variant tracing to a unique BC3F1 plant. The MABC lines were grown in spray trials at two locations throughout the United States with two replications at each location. Each line was planted in a paired, twelve foot plot. Each plot was sprayed at V3 with 0.75 lb a.e./acre of glyphosate and 1.0 lb a.e./acre dicamba followed by the same treatment at V6. Malformation was measured as the percentage of plants within a plot that were malformed. For each MABC, the data were averaged across variants, replications, and locations to place into one the following categories:

1. No malformation: No severe malformation detected

2. Malformation: >20% severe malformation

The data are presented in Table 4. The 6 MABC lines with a CCAATT or TTAATT haplotype all showed malformation whereas the 25 lines with the TTTTCC haplotype showed no malformation. These observations indicated that the presence of certain haplotypes at the dicamba tolerance locus in lines containing the DMO and RR2Y CP4 transgenes leads to malformation following dicamba treatment.

TABLE 4 Recurrent Parent Haplotypes and Malformation Phenotypes in Response to Dicamba Application. Recurrent Number of Response Parent Observations MG M0101742 M0205350 M0102027 to Dicamba AG3102 24 3 CC AA TT Malformation AG3603 30 3 CC AA TT Malformation BBL3606N0R 30 3 CC AA TT Malformation BL3307M2-D0RL 30 3 CC AA TT Malformation AG4903 36 4 TT AA TT Malformation AG4907 36 4 CC AA TT Malformation 260744-14 24 0 TT TT CC No Malformation AFL0506C0R 24 0 TT TT CC No Malformation AG0803 24 0 TT TT CC No Malformation AG0808 24 0 TT TT CC No Malformation 263619-24 24 1 TT TT CC No Malformation 4065735-51 24 1 TT TT CC No Malformation 5463213-25 36 1 TT TT CC No Malformation AG1002 24 1 TT TT CC No Malformation AG1403 24 1 TT TT CC No Malformation AG1406 24 1 TT TT CC No Malformation CSR1920 36 1 TT TT CC No Malformation 15733-79-59 30 2 TT TT CC No Malformation 5081541-27 12 2 TT TT CC No Malformation 5464705-06 30 2 TT TT CC No Malformation AG2110 36 2 TT TT CC No Malformation AG2606 29 2 TT TT CC No Malformation AG2909 30 2 TT TT CC No Malformation AG2921V 30 2 TT TT CC No Malformation AG3021V 30 3 TT TT CC No Malformation AG3803 30 3 TT TT CC No Malformation BOX2906H0R 30 3 TT TT CC No Malformation AG3803 30 3 TT TT CC No Malformation DFN3306B0R 24 3 TT TT CC No Malformation CSRS4782N 36 4 TT TT CC No Malformation GL4807A2-D0RN 36 4 TT TT CC No Malformation

Example 2: Haplotypes Associated with a Malformation and Sterility Phenotypes for MG 6 and 7 BC2F3:4 Populations Upon Herbicide Application

The effect of different haplotypes on response to dicamba and glyphosate were evaluated by comparing MG6-7 BC2F3:4 lines that contained a CP4 transgene that confers tolerance to glyphosate and a DMO transgene that confers tolerance to dicamba. The recurrent parent of the population was RP1 that had the CCAACC haplotype for markers NS0101742, NS0205350, and NS0102027 markers, respectively. The donor parent of the population was DP1 that had a TTTTCC haplotype. During the breeding process, the haplotype for each line at the dicamba tolerance locus was not known. Markers were used to select for the absence of the RR1 CP4 gene, and for the presence of the RR2Y CP4 gene and the Dicamba resistance DMO gene. Table 5 describes the breeding history for this material.

TABLE 5 Breeding History for Plant Material in Example 2. Gen. Season Year Location Breeding Activity Cross Winter 2007 Isabella, Puerto Rico Cross F1 Summer 2008 Isabella, Puerto Rico Backcross BC1F1 Winter 2008 Isabella, Puerto Rico Backcross BC2F1 Summer 2009 Isabella, Puerto Rico Bulk BC2F2 Winter 2009 Isabella, Puerto Rico Bulk BC2F3 Summer 2010 Mount Olive, NC Single plant selection BC2F4 Summer 2011 Mount Olive, NC Progeny row

A total of 360 BC2F3:4 lines were grown in Mount Olive, N.C. in 2011. The lines were grown in a single four foot rows with one replication. The lines were sprayed with 0.75 lb a.e./acre of glyphosate at V3 plant stage followed by the same rate of glyphosate at V6 plant stage plus 0.5 lb a.e./acre of dicamba.

Rating scales:

Malformation:

A rating of 1: Less than 10% of plants show malformation A rating of 2: 10-50% of plants show malformation A rating of 3: Greater than 50% of plants show malformation

Sterility:

A rating of 1: Less than 10% of plants show sterility A rating of 2: 10-50% of plants show sterility A rating of 3: Greater than 50% of plants show sterility

Table 6 shows the distribution of lines across the different rating classes.

TABLE 6 Malformation and Sterility Ratings for Soybean Populations. Malformation Rating 1 Sterility Rating Number of lines per rating class 2 3 1 154 18 3 2 58 61 52 3 2 2 10

The 51 lines that were rated “1” for malformation and “1” for sterility and the 62 lines that were rated “3” for malformation and “2” or “3” for sterility were genotyped for the three dicamba tolerance markers as shown in Table 7.

TABLE 7 Haplotypes Associated with a Malformation and Sterility Ratings for 51 Soybean Lines. Line Marker Haplotype Rating Number M0101742 M0205350 M0102027 Malformation Sterility 123 TT TT CC 1 1 124 CC AA CC 3 2 126 CC AA CC 3 2 128 TT TT CC 1 1 135 TT TT CC 1 1 136 CC AA CC 3 3 140 CC AA CC 3 2 141 TT TT CC 1 1 143 TT TT CC 1 1 144 CC AA CC 3 2 145 CC AA CC 3 2 149 CT AT CC 1 1 151 CC AA CC 3 2 154 CC AA CC 3 2 156 TT TT CC 1 1 157 CT AT CC 1 1 159 CC AA CC 3 2 160 CC AA CC 3 2 164 TT TT CC 1 1 171 CT AT CC 1 1 173 CC AA CC 3 2 184 CC AA CC 3 2 189 CC AA CC 3 2 193 CC AA CC 3 2 200 TT TT CC 1 1 201 CC AA CC 3 2 203 CC AA CC 3 2 204 CT AT CC 1 1 212 CC AA CC 3 2 213 TT TT CC 1 1 214 TT TT CC 1 1 216 CC AA CC 3 2 217 TT TT CC 1 1 218 TT TT CC 1 1 222 CT AT CC 3 2 223 CC AA CC 3 2 225 CT AT CC 1 1 228 TT TT CC 1 1 230 TT TT CC 1 1 234 CC AA CC 3 2 235 TT TT CC 1 1 243 CC AA CC 3 2 246 CC AA CC 1 1 251 CC AA CC 3 3 260 CC AA CC 3 3 261 CT AT CC 1 1 262 CT AT CC 3 3 264 CT AT CC 1 1 268 TT TT CC 1 1 274 CC AA CC 3 2 277 CC AA CC 3 2 280 CC AA CC 3 2 281 CC AA CC 3 2 290 CC AA CC 3 2 309 CC AA CC 1 1 314 CC AA CC 3 2 316 TT TT CC 1 1 317 CC AA CC 3 2 318 TT TT CC 1 1 319 TT TT CC 1 1 324 TT TT CC 1 1 329 CC AA CC 3 3 330 CC AA CC 1 1 331 CC AA CC 1 1 333 CC AA CC 3 2 334 CC AA CC 1 1 336 CC AA CC 3 2 337 CC AA CC 1 1 338 TT TT CC 1 1 340 CC AA CC 3 2 347 TT TT CC 1 1 348 TT TT CC 1 1 350 CT AT CC 3 2 352 CC AA CC 3 2 353 TT TT CC 1 1 362 CC AA CC 3 2 379 TT TT CC 1 1 380 TT TT CC 1 1 381 CC AA CC 3 2 382 CT AT CC 1 1 384 CC AA CC 3 2 385 CC AA CC 3 2 393 CT AT CC 1 1 396 TT TT CC 1 1 398 CC AA CC 3 2 399 CC AA CC 3 2 400 CC AA CC 3 2 408 CC AA CC 3 2 409 CC AA CC 3 2 411 TT TT CC 1 1 412 CC AA CC 3 2 417 TT TT CC 1 1 421 CC AA CC 3 2 433 CT AT CC 1 1 434 TT TT CC 1 1 435 TT TT CC 1 1 440 CC AA CC 3 2 446 CC AA CC 3 3 449 CC AA CC 3 3 452 TT TT CC 1 1 457 CC AA CC 3 3 460 CC AA CC 3 2 467 TT TT CC 1 1 468 CC AA CC 3 3 470 CC AA CC 3 2 488 CC AA CC 3 2 490 CC AA CC 3 2 494 CC AA CC 3 3 495 CC AA CC 3 2 500 CC AA CC 3 2 505 CC AA CC 3 2 510 TT TT CC 1 1 512 CC AA CC 1 1

The 34 lines with the TTTTCC haplotype had a rating of “1” for malformation and “1” for sterility as summarized in Table 8.

TABLE 8 Summary of Table 7. No. of Rating Marker Haplotype lines Malformation Sterility M0101742 M0205350 M0102027 34 1 1 TT TT CC 59 3 2 or 3 CC AA CC 7 1 1 10 1 1 CT AT CC 3 3 2 or 3

Out of the 66 lines with the CCAACC haplotype, 59 had a rating of “3” for malformation and a rating of “2” or “3” for sterility. The 13 lines heterozygous for the markers had a range of ratings for malformation and sterility.

These results support the observations that certain haplotypes at the dicamba tolerance locus in lines containing the RR2Y CP4 and DMO transgenes causes sterility from glyphosate and malformation from dicamba applications made at the V6 plant stage

Example 3. Haplotypes Associated with a Malformation and Sterility Phenotypes for MG 3 and 4 Populations Upon Herbicide Application in Fontezuela, Argentina

The effect of different haplotypes on response to glyphosate were evaluated by measuring observing sterility in MG3 to MG4 lines in glyphosate spray trials in Fontezuela, Argentina in 2012. The lines were from populations known to segregate for markers at the dicamba tolerance locus based on parental haplotypes. During the breeding process, the haplotype for each line at the dicamba tolerance locus was not known. Table 9 describes the breeding history for this material.

TABLE 9 Breeding History for Plant Material in Example 3. Gen. Season Year Location Breeding Activity Cross Winter or 2010 Isabella, Puerto Rico, Cross Summer or Galena, MD F1 Summer or 2010 Isabella, Puerto Rico Bulk Winter F2 Winter 2011 Kunia, HI Bulk F3 Summer 2011 Stonington, IL Single plant selection F4 Summer 2012 Fontezuela, Argentina Progeny row

Markers were used to select for the absence of the RR1 CP4 gene, and for the presence of the RR2Y CP4 gene and the Dicamba resistance (DMO) gene. A total of 1,083 F3:4 lines across six populations were planted in 4 foot single row plots. Remnant seed from each line was used for genotyping the lines across two markers at the dicamba tolerance locus. The lines were sprayed with 1.125 lb a.e./acre of glyphosate at V3 plant stage followed by the same rate applied at V6. Sterility ratings were taken at maturity.

A rating of 1: Less than 10% of plants show sterility A rating of 2: 10-50% of plants show sterility A rating of 3: Greater than 50% of plants show sterility

The sterility ratings by haplotype class are shown in Table 10.

TABLE 10 Haplotypes Associated with sterility ratings for Selected Soybean Populations. Total No. lines per Marker Haplotype No. of rating class Population M0101742 M0205350 Lines 1 2 3 POP1 TT TT 42 42 CC AA 27 10 17 CT AT 25 1 23 1 POP2 TT TT 83 83 TT AA 23 8 15 TT AT 36 11 25 POP3 TT TT 81 81 CC AA 89 24 65 CT AT 40 4 27 9 POP4 TT TT 69 69 CC AA 43 1 2 40 CT AT 39 1 27 11 TT TT 75 75 TT AA 62 3 59 TT AT 44 44 POP5 TT TT 185 185 CC AA 66 2 64 CT AT 54 4 47 3 Across TT TT 535 535 0 0 populations CC AA 225 1 38 186 (POP1-5) TT AA 85 0 11 74 CT AT 158 10 124 24 TT AT 80 0 55 25

Lines with the TTTT haplotype for markers M0101742 and M0205350 did not show sterility across all populations. Nearly all lines with a CCAA or TTAA haplotype had at least 10% of plants that showed sterility. It is not unexpected that some plants in these lines did no show sterility as some variation in the spray application or other environmental variations can influence the expression of sterility. Lines genotyped as CTAT or TTAT were segregating at the dicamba tolerance locus and progeny from these lines showed a range of response for sterility.

These results support the observations that certain haplotypes at the dicamba tolerance locus in lines containing the RR2Y CP4 and DMO transgenes causes sterility from glyphosate applications made at the V6 plant stage.

Example 4: Exemplary Marker Assays for Detecting Polymorphisms

In one embodiment, the detection of polymorphic sites in a sample of DNA, RNA, or cDNA may be facilitated through the use of nucleic acid amplification methods. Such methods specifically increase the concentration of polynucleotides that span the polymorphic site, or include that site and sequences located either distal or proximal to it. Such amplified molecules can be readily detected by gel electrophoresis, fluorescence detection methods, or other means. Exemplary primers and probes for amplifying and detecting genomic regions associated with a dicamba tolerance phenotype are given in Table 11.

TABLE 11 Exemplary Assays for Detecting Polymorphisms SEQ ID NO SEQ ID NO Marker or Marker SNP Forward Reverse SEQ ID NO SEQ ID NO Locus Name SEQ NO ID: Position Primer Primer Probe 1 Probe 2 asmbl_11856 1 TC122822 2 BI967232 3 M0205928 4 M0101742³ 5 1206 24 25 26 27 M0129138 6 218 28 29 30 31 BU551345 7 M0114388 8 502 32 33 34 35 BU551363 9 M0205350⁴ 10 148 36 37 38 39 M0102027⁵ 11 349 40 41 42 43 BU765955 12 M0093116 13 183 44 45 46 47 M0129925 14 328 48 49 50 51 M0205537 15 M0202715 16 M0206286 17 M0206054 18 M0205375 19 NGMAX008197032 52 201 53 54 55 56

Example 5: Oligonucleotide Probes Useful for Detecting Polymorphisms by Single Base Extension Methods

Oligonucleotides can also be used to detect or type the polymorphisms disclosed herein by single base extension (SBE)-based SNP detection methods. Exemplary oligonucleotides for use in SBE-based SNP detection are provided in Table 12. SBE methods are based on extension of a nucleotide primer that is hybridized to sequences adjacent to a polymorphism to incorporate a detectable nucleotide residue upon extension of the primer. It is also anticipated that the SBE method can use three synthetic oligonucleotides. Two of the oligonucleotides serve as PCR primers and are complementary to the sequence of the locus which flanks a region containing the polymorphism to be assayed. Exemplary extension primers that can be used to type polymorphisms disclosed in this invention are provided in Table 12 in the column labeled “Probe (SBE)”. Following amplification of the region containing the polymorphism, the PCR product is hybridized with an extension primer which anneals to the amplified DNA adjacent to the polymorphism. DNA polymerase and two differentially labeled dideoxynucleoside triphosphates are then provided. If the polymorphism is present on the template, one of the labeled dideoxynucleoside triphosphates can be added to the primer in a single base chain extension. The allele present is then inferred by determining which of the two differential labels was added to the extension primer. Homozygous samples will result in only one of the two labeled bases being incorporated and thus only one of the two labels will be detected. Heterozygous samples have both alleles present, and will thus direct incorporation of both labels (into different molecules of the extension primer) and thus both labels will be detected. Exemplary forward and reverse SBE probes are provided in Table 12.

TABLE 12 Exemplary SBE Probes for Detecting Polymorphisms Marker Probe Marker or Locus (SEQ ID SNP (SEQ ID Name NO) Position Probe (SBE) NO) M0101742  5 1206 TGACTAGCATGTATCTAT 26 ATGACTAACATGTATCTAT 27 M0129138  6  218 TGTGTCCTATATGATCTT 30 TGTCCTGTATGATCTTA 31 M0114388  8  502 AGTTGGGCTATGCAA 34 TGGGCTGTGCAAGTA 35 M0205350 10  148 AGTTTACACTTACAAATATT 38 AGAGTTTACACTTACATATATT 39 M0102027 11  349 ACCCCCCTTTTTT 42 ATTTTAACCCCCTTTTT 43 M0093116 13  183 CCAACACCAAACTA 46 CAACACCAAACAAA 47 M0129925 14  328 AGTAGTAGCTAGTGAAATA 50 AGCTAGTCAAATATTT 51 NGMAX008197032 52  201 TTGACAGCCTCTGGATAT 55 ACAGCCTCCGGATAT 56

Example 6: Haplotypes Associated with a Malformation and Sterility Phenotypes in MG 4 Populations Upon Herbicide Application

The effect of different haplotypes on response to dicamba and glyphosate were evaluated by comparing genetically similar lines that contained a CP4 transgene that confers tolerance to glyphosate and a DMO transgene that confers tolerance to dicamba. In 2010, two plants from each of fourteen BC1F2:4 lines, or families, across five backcross populations were harvested individually to develop pairs of BC1F4:6 lines from each family. The haplotype of each recurrent parent for each backcross population is shown in Table 13.

TABLE 13 Haplotypes Associated With Listed Recurrent Parent. Backcross Recurrent parent haplotype population Recurrent parent M0101742 M0205350 M0102027 1 CBL3606Q0R CC AA TT 2 AG4005 CC AA TT 3 CP4408A3-C0RN CC AA TT 4 AG4907 CC AA TT 5 AG4630 TT AA TT

The donor parent for each population was A3244-RR2Y/A3525-DT that had a TTTTCC haplotype for markers M0101742, M0205350, and M0102027 markers, respectively. During the breeding process, the haplotype for each line at the dicamba tolerance locus was not known. Markers were used to select for the absence of the RR1 CP4 gene, and for the presence of the RR2Y CP4 gene and the Dicamba DMO gene. The BC1F4:5 rows were sprayed with glyphosate at the V6 plant growth stage in Quillota Chile where a pair of lines per family were rated for sterility to glyphosate. Table 14 describes the breeding history for this plant material.

TABLE 14 Breeding History for Plant Material in Example 6. Gen. Season Year Location Breeding Activity Cross Winter 2007 Isabella, Puerto Rico Cross F1 Summer 2008 Isabella, Puerto Rico Backcross BC1F1 Winter 2008 Isabella, Puerto Rico Bulk BC1F2 Summer 2009 Evansville, IN, Single plant selection Stonington, IL, or Galena, MD BC1F2: Summer 2010 Fontezuela, Argentina Progeny row 3 BC1F2: Summer 2010 Evansville, IN, Single plant selection 4 Stonington, IL, or Galena, MD BC1F4: Summer 2011 Quillota, Chile Progeny row 5 BC1F4: Summer 2011 Evansville, IN, Spray trials 6 Stonington, IL, Galena, MD, and Stuttgart, AR

DNA was extracted from each line to generate haplotypes across three markers at the dicamba tolerance locus. The lines were evaluated across four locations in the United States (Stuttgart, Ark.; Stonington Ill.; Evansville, Ind.; and Galena, Md.) in 2011. At each location the lines were grown in four to five foot single-row plots replicated two times and one of seven different herbicide treatments were applied at different plant growth stages (V3 or V6) as described in Table 15.

TABLE 15 Herbicide Treatments (Glyphosate and Dicamba) and Concentrations Applied at Plant Growth Stages (V3 or V6). Herbicide Treatment Glyphosate Dicamba Glyphosate Dicamba Number V3 V3 V6 V6 1 none none none none 2 0.75 lb none none none a.e./acre 3 0.75 lb none 1.5 lb none a.e./acre a.e./acre 4 none 1.0 lb none none a.e./acre 5 none 1.0 lb none 1.0 lb a.e./acre a.e./acre 6 0.75 lb 1.0 lb none none a.e./acre a.e./acre 7 0.75 lb 1.0 lb 1.5 lb 1.0 lb a.e./acre a.e./acre a.e./acre a.e./acre

Rating Scale:

Malformation to dicamba was rated by the percentage of plants showing malformation Sterility to glyphosate was rated as: A rating scale of 1: Less than 10% of plants show sterility A rating scale of 2: Less than 10-50% of plants show sterility A rating scale 3: Greater than 50% of plants show sterility

Data were averaged across replications and locations to place into following classes as described in Table 16.

TABLE 16 Haplotypes Associated with a Malformation and Sterility Phenotypes in Soybean Sister Line Pedigrees in Response to Herbicide Treatment Protocols. Reaction to Glyphosate Treatment No. Backcross in Quillota, Marker haplotype 1 2 3 4 5 6 7 Population Family Chile M0101742 M0205350 M0102027 Response 1 1.1 N TT TT CC N N N N N N N S CC AA TT N N S N M N M/S 2 2.1 S CC AA TT N N S N M N M/S S CC AA TT N N S N M N M/S 3 3.1 N TT TT CC N N N N N N N S CC AA TT N N S N M N M/S 4 4.1 S CC AA TT N N S N M N M/S N TT TT CC N N N N N N N 4.2 S CC AA TT N N S N M N M/S S CC AA TT N N S N M N M/S 4.3 S CC AA TT N N S N M N M/S S CC AA TT N N S N M N M/S 4.4 S CC AA TT N N S N M N M/S S CC AA TT N N S N M N M/S 4.5 S CC AA TT N N S N M N M/S S CC AA TT N N S N M N M/S 4.6 N TT TT CC N N N N N N N N TT TT CC N N N N N N N 4.7 N TT TT CC N N N N N N N S CC AA TT N N S N M N M/S 4.8 S CC AA TT N N S N M N M/S S CC AA TT N N S N M N M/S 4.9 S CC AA TT N N S N M N M/S S CC AA TT N N S N M N M/S 4.10 S CC AA TT N N S N M N M/S S CC AA TT N N S N M N M/S 5 5.1 S TT AA TT N N S N M N M/S N TT TT CC N N N N N N N N = normal response to treatment: Average </=1.25 sterility and/or </=30% malformation S = sterility to a glyphosate treatment: Average >1.25 M = malformation to a dicamba treatment >30% malformation M/S = malformation dicamba and sterility to glyphosate in a combination treatment

Example 7: Selection for Absence of Sterility to Glyphosate Application and Recovery of the Favorable Haplotype

As described in Example 3, marker haplotypes corresponded to reaction to glyphosate application. There were several populations grown in Fontezuela, Argentina where plants sprayed with glyphosate were selected for reproductive tolerance to glyphosate in the absence of haplotype information on each line. Table 17 describes four populations that segregated for the haplotype based on parental haplotypes. Table 18 describes the number of lines grown and the number of lines selected.

TABLE 17 Four soybean populations that segregated for the preferred haplotype based on parental haplotypes. Parent 1 Parent 2 NGMAX-008197032 M-0205350 NGMAX-008197032 M-0205350 Population Origin (SEQ ID NO: 52) (SEQ ID NO: 10) (SEQ ID NO: 52) (SEQ ID NO: 10) 1 AG4031/AG3803- GG AA AA TT T0BAH 2 AG4130/AG4907- AA TT GG AA T0BAH 3 BL3510A9- AA TT GG AA D0AAC/AG4907- T0BAH 4 EI4409C3- GG AA AA TT D0YN/GL4807A2- D0RN- T0BAH

TABLE 18 Number of soybean lines grown and the number of soybean lines selected. Population No. Lines Grown No. Lines Selected 1 165 2 2 92 9 3 200 10 4 260 25 Total 717 46

The 46 selected lines were subsequently genotyped and found to possess the favorable AATT haplotype. In addition, the lines were grown at Stonington, Ill. in 2012 and evaluated for herbicide response. The lines were sprayed with 1.0 lb a.e/acre dicamba and 1.5 lb a.e/acre glyphosate at the V6 plant stage and did not show malformation to dicamba or sterility to glyphosate. These results further support the ability to use glyphosate selection as a means to recover the favorable haplotype and tolerance to both glyphosate and dicamba.

Example 8. Comparison of Dicamba Tolerance in Different Haplotypes Containing a Dicamba Resistance Conferring Transgene

The effect of different haplotypes on response to dicamba was evaluated by comparing F2 families that contained the DMO transgene for dicamba resistance, but that lacked the CP4 transgene for glyphosate resistance. F2 plants across six different populations that were growing in Kunia, Hi. in 2012 were tissue sampled and genotyped for the CP4 and DMO transgenes and for markers NGMAX008197032 and M0205350. F2 plants that were fixed homozygous for the presence of DMO and absence of CP4 and that were fixed homozygous for a haplotype class were selected and harvested individually to create families. Table 19 shows the number of F2 families per haplotype class. The F2 families were evaluated for tolerance to dicamba in a greenhouse environment.

TABLE 19 Six soybean populations that segregated for the preferred haplotype based on parental haplotypes. Haplotype Class AATT¹ GGAA² POP ORIGIN Number of F2 plants 5 A3525-A3244-BAH/A3431 8 10 6 DKB31-51/A3525-A3244-BAH 3 3 8 AG4903/A3525-A3244-BAH 0 3 9 AG4907/A3525-A3244-BAH 2 2 12 AG4903/(AG4903*2/A3525-A3244-BAH) 0 5 13 AG4907/GL4911A9-B0BAH 0 5 ¹An “AA” allele for NGMAX-008197032 (SEQ ID NO: 52) and a “TT” allele for M020535(SEQ ID NO: 10) (i.e. “favorable” haplotype for dicamba tolerance). ²A “GG” allele for NGMAX-008197032 (SEQ ID NO: 52) and an “AA” allele for M020535(SEQ ID NO: 10) (i.e. “unfavorable” haplotype for dicamba tolerance).

A comparison of dicamba tolerance in the plants from segregating populations of Table 19 having various “favorable” or “unfavorable” haplotypes of the indicated parental germplasm is provided in FIG. 1. Plants having the favorable haplotypes (i.e. an “AA” allele for NGMAX-008197032 (SEQ ID NO:52) and a “TT” allele for M020535(SEQ ID NO: 10) showed consistently low dicamba injury (FIG. 1).

Example 9. Selection of Favorable Dicamba Tolerance Haplotypes with One or Two Spray Treatments

A comparison of selections of favorable and unfavorable dicamba tolerance haplotypes based on either one or two spray treatments was made. Transgenic soybean plants containing a dicamba resistance conferring transgene and various favorable or unfavorable haplotypes were treated with dicamba at a rate of 1 pound/acre at either: (a) the V3 and V6 stages; or (b) the V6 stage only. The results of this comparison are shown in FIG. 2. Selection of favorable haplotypes by using a single spray at V6 (dicamba 1 lb/a) was found to be as effective as selections with the combination of aV3 and a V6 spray treatment.

Having illustrated and described the principles of the present invention, it should be apparent to persons skilled in the art that the invention can be modified in arrangement and detail without departing from such principles.

Although the materials and methods of this invention have been described in terms of various embodiments and illustrative examples, it will be apparent to those of skill in the art that variations can be applied to the materials and methods described herein without departing from the concept, spirit and scope of the invention. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

TABLE 2 of the Specification Locus/Display Start End ADDITIONAL LOCUS Name (1) Source (2) Base (3) Base (4) INFORMATION (5) asmbl_11856 Vigna _(—) unguiculata 16426 16586 SEQ ID NO: 1 TA2790_3886 Phaseolus _(—) coccineus _(—) 16423 17393 ADP-ribosylation factor release_2 [Vigna unguiculata (Cowpea)] TA43459_3847 Glycine _(—) max_release_2 16434 18055 ADP-ribosylation factor 1 [Oryza sativa (Rice)] TC276541 GMGI.071508 16434 18076 UniRef100_P36397 Cluster: ADP-ribosylation factor 1; n = 1; Arabidopsis thaliana|Rep: ADP- ribosylation factor 1 - Arabidopsis thaliana (Mouse-ear cress) = partial (38%) CD392203 Glycine _(—) max_release_2 16216 18687 ADP-ribosylation factor [Glycine max (Soybean)] BQ610865 Glycine _(—) max_release_2 16327 18667 ADP-ribosylation factor 1 [Oryza sativa (Rice)] EH046324 Arachis _(—) stenosperma _(—) 16405 18745 Cluster: ADP-ribosylation release_5 factor 1, n = 1, Arabidopsis thaliana|Rep: ADP- ribosylation factor 1 - Arabidopsis thaliana (Mouse-ear cress) AW202311 Glycine _(—) max_release_2 16378 19070 ADP-ribosylation factor [Glycine max (Soybean)] TC242702 GMGI.071508 16234 20195 UniRef100_Q38JU3 Cluster: ADP ribosylation factor 002; n = 2; core eudicotyledons|Rep: ADP ribosylation factor 002 - Daucus carota (Carrot) = complete BI321678 Glycine _(—) max_release_2 17384 19066 ADP-ribosylation factor [Zea mays (Maize)] AW348317 Glycine _(—) max_release_2 16355 20097 ADP-ribosylation factor [Glycine max (Soybean)] EH042959 Arachis _(—) stenosperma _(—) 16401 20182 Cluster: ADP-ribosylation release_5 factor 1, n = 2, Medicago|Rep: ADP- ribosylation factor 1 - Medicago truncatula (Barrel medic) TC20337 LJGI.070108 16420 20191 UniRef100_Q5QQ33 Cluster: ADP-ribosylation factor 1, n = 2, Medicago|Rep: ADP- ribosylation factor 1 - Medicago truncatula (Barrel medic), complete EH047563 Arachis _(—) stenosperma _(—) 16430 20182 Cluster: ADP-ribosylation release_5 factor 1, n = 2, Medicago|Rep: ADP- ribosylation factor 1 - Medicago truncatula (Barrel medic) TA2789_3886 Phaseolus _(—) coccineus _(—) 16436 20196 ADP-ribosylation factor 1- release_2 like protein [Solanum tuberosum (Potato)] TA43462_3847 Glycine _(—) max _(—) 16229 20438 ADP-ribosylation factor release_2 [Medicago sativa (Alfalfa)] TA1120_34305 Lotus _(—) japonicus _(—) 16522 20191 ADP-ribosylation factor release_1 [Medicago sativa (Alfalfa)] TA2306_3848 Glycine _(—) soja _(—) 16442 20440 ADP-ribosylation factor release_2 [Medicago sativa (Alfalfa)] TC273941 GMGI.071508 16426 20464 homologue to UniRef100_Q38JU3 Cluster: ADP ribosylation factor 002; n = 2; core eudicotyledons|Rep: ADP ribosylation factor 002 - Daucus carota (Carrot) = complete TC238119 GMGI.071508 16455 20449 UniRef100_Q38JU3 Cluster: ADP ribosylation factor 002; n = 2; core eudicotyledons|Rep: ADP ribosylation factor 002 - Daucus carota (Carrot) = complete EG373880 Arachis _(—) hypogaea _(—) 17101 20182 Cluster: ADP-ribosylation release_5 factor 1, n = 2, Medicago|Rep: ADP- ribosylation factor 1 - Medicago truncatula (Barrel medic) BF066818 Glycine _(—) max_release_2 17081 20378 ADP-ribosylation factor 1 [Populus tomentosa] BF596154 Glycine _(—) max_release_2 17083 20397 ADP-ribosylation factor [Hyacinthus orientalis (Common hyacinth)] AW760997 Glycine _(—) max_release_2 17116 20397 ADP-ribosylation factor [Hyacinthus orientalis (Common hyacinth)] BF424079 Glycine _(—) max_release_2 17112 20417 ADP-ribosylation factor [Hyacinthus orientalis (Common hyacinth)] AW596022 Glycine _(—) max_release_2 17121 20415 ADP-ribosylation factor 1 [Populus tomentosa] TA43446_3847 Glycine _(—) max_release_2 17106 20436 ADP-ribosylation factor [Hyacinthus orientalis (Common hyacinth)] TA43455_3847 Glycine _(—) max_release_2 17125 20452 ADP-ribosylation factor [Hyacinthus orientalis (Common hyacinth)] BW595867 Lotus _(—) japonicus _(—) 17418 20191 ADP-ribosylation factor release_1 [Hyacinthus orientalis (Common hyacinth)] AW507598 Glycine _(—) max_release_2 17343 20437 ADP-ribosylation factor [Hyacinthus orientalis (Common hyacinth)] TA43447_3847 Glycine _(—) max_release_2 17343 20445 ADP-ribosylation factor [Hyacinthus orientalis (Common hyacinth)] TA43448_3847 Glycine _(—) max_release_2 17355 20438 ADP-ribosylation factor 1 [Populus tomentosa] AW596189 Glycine _(—) max_release_2 17358 20442 ADP-ribosylation factor 1 [Populus tomentosa] BI469983 Glycine _(—) max_release_2 17410 20438 ADP-ribosylation factor 1 [Populus tomentosa] AW472058 Glycine _(—) max_release_2 18655 20160 ADP-ribosylation factor 1 [Daucus carota (Carrot)] CB063805 Glycine _(—) max_release_2 18623 20432 ADP-ribosylation factor 1 [Populus tomentosa] BM891090 GMGI.071508 18995 20429 homologue to UniRef100_A7PRL9 Cluster: Chromosome chr14 scaffold_27 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_27 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (42%) BM731935 Glycine _(—) max_release_2 19949 20444 ADP-ribosylation factor 1 [Populus tomentosa] AW695591 MTGI.071708 30054 31388 similar to UniRef100_Q40542 Cluster: NPK2, n = 1, Nicotiana tabacum|Rep: NPK2 - Nicotiana tabacum (Common tobacco), partial (35%) TC130040 MTGI.071708 30054 31482 similar to UniRef100_A7PM42 Cluster: Chromosome chr14 scaffold_21, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr14 scaffold_21, whole genome shotgun sequence - Vitis vinifera (Grape), partial (30%) TC122822 MTGI.071708 30054 34162 Protein kinase, Nuclear transport factor 2. SEQ ID NO: 2 Pvcon9203 Phaseolus _(—) vulgaris 31194 34247 UniRef100_A7PM42 Chromosome chr14 scaffold_21, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7PM42_VITVI E−0 TA66103_3847 Glycine _(—) max_release_2 31879 34559 Protein kinase; Nuclear transport factor 2 [Medicago truncatula (Barrel medic)] CA801261 GMGI.071508 33896 34304 similar to UniRef100_Q40542 Cluster: NPK2; n = 1; Nicotiana tabacum|Rep: NPK2 - Nicotiana tabacum (Common tobacco) = partial (16%) TC120073 MTGI.071708 35367 38178 Glycoside hydrolase, family 28 NP004759 GMGI.071508 34976 39622 GB|AF128266.1|AAD46483.1 polygalacturonase PG1 AF128266 Glycine _(—) max_release_2 34980 39622 Polygalacturonase PG1 [Glycine max (Soybean)] TA69799_3847 Glycine _(—) max_release_2 58988 65870 Ubiquitin-associated [Medicago truncatula (Barrel medic)] TA7619_47247 Lotus _(—) corniculatus _(—) 63855 65940 Putative DNA cytosine release_1 methyltransferase Zmet3 related cluster TA8711_34305 Lotus _(—) japonicus _(—) 63855 65940 UBA-like [Medicago release_1 truncatula (Barrel medic)] TC34762 LJGI.070108 65619 65940 NA Pvcon5587 Phaseolus _(—) vulgaris 65216 67090 UniRef100_A7PM76 Chromosome chr14 scaffold_21, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7PM76_VITVI E−0 TA5046_3885 Phaseolus _(—) vulgaris _(—) 65808 67002 UBA-like [Medicago release_2 truncatula (Barrel medic)] asmbl_11857 Vigna _(—) unguiculata 65951 67042 NA TA58707_3847 Glycine _(—) max_release_2 66006 67253 UBA-like [Medicago truncatula (Barrel medic)] TC241193 GMGI.071508 66006 67253 similar to UniRef100_A7PM76 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (38%) BI967232 Glycine _(—) max_release_2 66170 67203 UBA-like [Medicago truncatula (Barrel medic)]. SEQ ID NO: 3 AV417590 LJGI.070108 66745 67090 similar to UniRef100_A7PM76 Cluster: Chromosome chr14 scaffold_21, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr14 scaffold_21, whole genome shotgun sequence - Vitis vinifera (Grape), partial (19%) AV768315 Lotus _(—) japonicus _(—) 66699 67155 UBA-like [Medicago release_1 truncatula (Barrel medic)] TC32114 LJGI.070108 66699 67275 similar to UniRef100_Q76KU6 Cluster: DNA methyltransferase, n = 1, Nicotiana tabacum|Rep: DNA methyltransferase - Nicotiana tabacum (Common tobacco), partial (20%) TA1535_34305 Lotus _(—) japonicus _(—) 66745 67277 UBA-like [Medicago release_1 truncatula (Barrel medic)] TA2793_47247 Lotus _(—) corniculatus _(—) 66745 67277 DNA methyltransferase release_1 related cluster AV768911 Lotus _(—) japonicus _(—) 66943 67155 Ubiquitin-associated release_1 [Medicago truncatula (Barrel medic)] CB540531 Phaseolus _(—) vulgaris 73267 73561 UniRef100_A7PM74 Chromosome chr14 scaffold_21, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7PM74_VITVI 5.00E−27 BE347690 GMGI.071508 73509 73770 similar to UniRef100_Q5VQL1-2 Cluster: Isoform 2 of Q5VQL1; n = 1; Oryza sativa Japonica Group|Rep: Isoform 2 of Q5VQL1 - Oryza sativa subsp. japonica (Rice) = partial (5%) BE347690 Glycine _(—) max_release_2 73509 73822 WW/Rsp5/WWP; Helicase = C-terminal [Medicago truncatula (Barrel medic)] BE608496 GMGI.071508 73444 73947 similar to UniRef100_A7PM74 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (16%) AI416763 GMGI.071508 74073 74520 similar to UniRef100_Q9SP26 Cluster: P72 DEAD box protein; n = 1; Pisum sativum|Rep: P72 DEAD box protein - Pisum sativum (Garden pea) = partial (16%) AI416763 Glycine _(—) max_release_2 74073 74743 ATP-dependent RNA helicase-like protein DB10 [Nicotiana sylvestris (Wood tobacco)] BW615083 LJGI.070108 74256 74855 similar to UniRef100_A7PM74 Cluster: Chromosome chr14 scaffold_21, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr14 scaffold_21, whole genome shotgun sequence - Vitis vinifera (Grape), partial (24%) TA8332_34305 Lotus _(—) japonicus _(—) 74256 75446 WW/Rsp5/WWP, Helicase, release_1 C-terminal [Medicago truncatula (Barrel medic)] TC27807 LJGI.070108 74343 75446 similar to UniRef100_Q9SP26 Cluster: P72 DEAD box protein, n = 1, Pisum sativum|Rep: P72 DEAD box protein - Pisum sativum (Garden pea), partial (34%) asmbl_11858 Vigna _(—) unguiculata 75228 75500 NA TA60825_3847 Glycine _(—) max_release_2 74963 75981 P72 DEAD box protein [Pisum sativum (Garden pea)] TC249436 GMGI.071508 74985 75966 similar to UniRef100_Q9SP26 Cluster: P72 DEAD box protein; n = 1; Pisum sativum|Rep: P72 DEAD box protein - Pisum sativum (Garden pea) = partial (12%) TC269249 GMGI.071508 86882 87576 similar to UniRef100_A7PM72 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (42%) TA64136_3847 Glycine _(—) max_release_2 86882 89066 Putative phosphate/phosphoenolpyruvate translocator [Arabidopsis thaliana (Mouse-ear cress)] CO982132 Glycine _(—) max_release_2 87225 91497 Phosphate/phosphoenolpyruvate translocator [Nicotiana tabacum (Common tobacco)] TC274531 GMGI.071508 87225 91497 similar to UniRef100_A4UTS3 Cluster: Chloroplast phosphoenolpyruvate/ phosphate translocator; n = 1; Pisum sativum|Rep: Chloroplast phosphoenolpyruvate/ phosphate translocator - Pisum sativum (Garden pea) = partial (53%) Pvcon2802 Phaseolus _(—) vulgaris 87119 92616 UniRef100_A9PD12 Putative uncharacterized protein n = 1 Tax = Populus trichocarpa RepID = A9PD12_POPTR 1.00E−121 TA4406_3885 Phaseolus _(—) vulgaris _(—) 89055 92616 Phosphate/phosphoenolpyruvate release_2 translocator [Nicotiana tabacum (Common tobacco)] TA74766_3847 Glycine _(—) max_release_2 91397 92725 Phosphoenolpyruvate/ phosphate translocator [Mesembryanthemum crystallinum (Common ice plant)] TC265023 GMGI.071508 91686 92725 similar to UniRef100_A7PM71 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (15%) M0205928 SEQ. LISTING 92718 92334 SEQ ID NO: 4 BG406195 GMGI.071508 107039 107366 BG406195 Glycine _(—) max_release_2 107039 107375 NA M0101742 SEQ. LISTING 112189 113483 SEQ ID NO: 5 BG550728 GMGI.071508 112663 113757 weakly similar to UniRef100_A7PM60 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (13%) BG550728 Glycine _(—) max_release_2 112663 113867 Receptor-like serine/threonine kinase [Arabidopsis thaliana (Mouse-ear cress)] CV535605 Phaseolus _(—) vulgaris 112548 113982 UniRef100_A7PM60 Chromosome chr14 scaffold_21, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7PM60_VITVI 9.00E−79 M0129138 SEQ. LISTING 114532 113494 SEQ ID NO: 6 BU551345 Glycine _(—) max_release_2 115956 116339 SEQ ID NO: 7 TA58315_3847 Glycine _(—) max_release_2 118318 120087 NA TC236438 GMGI.071508 118318 120087 NA BE611751 Glycine _(—) max_release_2 119165 119645 NA BE611751 GMGI.071508 119229 119645 NA TA70371_3847 Glycine _(—) max_release_2 137417 137864 Hypothetical protein [Medicago truncatula (Barrel medic)] TC267549 GMGI.071508 137417 137864 similar to UniRef100_Q9FI64 Cluster: Genomic DNA = chromosome 5 = TAC clone: K21I16; n = 1; Arabidopsis thaliana|Rep: Genomic DNA = chromosome 5 = TAC clone: K21I16 - Arabidopsis thaliana (Mouse-ear cress) = partial (43%) BG156330 GMGI.071508 155872 156903 similar to UniRef100_A7PM41 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 2; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (23%) BG156330 Glycine _(—) max_release_2 155872 157058 WD40-like [Medicago truncatula (Barrel medic)] Pvcon10326 Phaseolus _(—) vulgaris 155691 157835 UniRef100_A7PM41 Chromosome chr14 scaffold_21, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7PM41_VITVI 3.00E−93 CD397113 Glycine _(—) max_release_2 157474 157813 NA TA12653_34305 Lotus _(—) japonicus _(—) 159489 161341 NADP-specific isocitrate release_1 dehydrogenase [Lupinus albus (White lupin)] TC27381 LJGI.070108 159489 161341 similar to UniRef100_Q7Y0W7 Cluster: NADP-specific isocitrate dehydrogenase, n = 1, Lupinus albus|Rep: NADP-specific isocitrate dehydrogenase - Lupinus albus (White lupin), partial (25%) DT084057 Glycine _(—) soja _(—) 161638 162192 NADP-specific isocitrate release_2 dehydrogenase [Lupinus albus (White lupin)] BE661051 Glycine _(—) max_release_2 170271 172034 Cyclin-like F-box [Medicago truncatula (Barrel medic)] TA11305_34305 Lotus _(—) japonicus _(—) 170700 172307 Cyclin-like F-box release_1 [Medicago truncatula (Barrel medic)] TC34049 LJGI.070108 170700 172307 similar to UniRef100_A7PF14 Cluster: Chromosome chr11 scaffold_13, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr11 scaffold_13, whole genome shotgun sequence - Vitis vinifera (Grape), partial (32%) NP7256876 MTGI.071708 171929 173188 GB|AC157983.16|ABE865 10.1 Cyclin-like F-box TA68495_3847 Glycine _(—) max_release_2 194920 195696 Oleosin [Sesamum indicum (Oriental sesame) (Gingelly)] TC265354 GMGI.071508 194920 195696 weakly similar to UniRef100_P29530 Cluster: P24 oleosin isoform A; n = 1; Glycine max|Rep: P24 oleosin isoform A - Glycine max (Soybean) = partial (40%) BE658264 Glycine _(—) max_release_2 195176 195925 Oleosin [Sesamum indicum (Oriental sesame) (Gingelly)] CV539661 Phaseolus _(—) vulgaris 217885 218101 No significant hit (e−20) CA912681 Phaseolus _(—) coccineus _(—) 220374 220748 Arabidopsis thaliana release_2 genomic DNA, chromosome 3, P1 clone: MGF10 [Arabidopsis thaliana (Mouse-ear cress)] CA785107 Glycine _(—) soja _(—) 221393 221885 NA release_2 TC276537 GMGI.071508 221407 222104 weakly similar to UniRef100_Q4RYK7 Cluster: Chromosome 3 SCAF14975 = whole genome shotgun sequence; n = 1; Tetraodon nigroviridis|Rep: Chromosome 3 SCAF14975 = whole genome shotgun sequence - Tetraodon nigroviridis (Green puffer) = partial (21%) TA71044_3847 Glycine _(—) max_release_2 221407 222133 NA CD406643 Glycine _(—) max_release_2 222113 222297 NA AV416316 LJGI.070108 223773 223869 similar to UniRef100_A7PM35 Cluster: Chromosome chr14 scaffold_21, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr14 scaffold_21, whole genome shotgun sequence - Vitis vinifera (Grape), partial (9%) EC911350 Phaseolus _(—) vulgaris 224587 225958 UniRef100_A5C233 Putative uncharacterized protein n = 1 Tax = Vitis vinifera RepID = A5C233_VITVI 3.00E−77 BU760697 GMGI.071508 224857 225965 similar to UniRef100_A7PM35 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (22%) BU760697 Glycine _(—) max_release_2 224857 226145 Protein At5g19130 [Arabidopsis thaliana (Mouse-ear cress)] TC119982 MTGI.071708 224248 226812 Gaa1-like, GPI transamidase component CV541515 Phaseolus _(—) vulgaris 225934 226374 UniRef100_A7PM35 Chromosome chr14 scaffold_21, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7PM35_VITVI 2.00E−34 TA76349_3847 Glycine _(—) max_release_2 226118 226768 Protein At5g19130 [Arabidopsis thaliana (Mouse-ear cress)] TA12045_47247 Lotus _(—) corniculatus _(—) 226354 226789 GPAA1-like protein related release_1 cluster TA13675_34305 Lotus _(—) japonicus _(—) 226354 226789 Protein At5g19130 release_1 [Arabidopsis thaliana (Mouse-ear cress)] TC29330 LJGI.070108 226354 226789 similar to UniRef100_A7PM35 Cluster: Chromosome chr14 scaffold_21, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr14 scaffold_21, whole genome shotgun sequence - Vitis vinifera (Grape), partial (13%) NP7254537 MTGI.071708 233411 237212 GB|AC152349.11|ABP03404.1 Protein of unknown function DUF266, plant EH256962 GMGI.071508 235306 237649 similar to UniRef100_A7PM54 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (33%) CX708677 Glycine _(—) max_release_2 247269 248145 NA BW599077 LJGI.070108 255475 261945 similar to UniRef100_A7QD90 Cluster: Peptidyl-prolyl cis- trans isomerase, n = 1, Vitis vinifera|Rep: Peptidyl- prolyl cis-trans isomerase - Vitis vinifera (Grape), partial (18%) BW625918 LJGI.070108 257810 262980 similar to UniRef100_Q93YQ8 Cluster: Peptidyl-prolyl cis- trans isomerase, n = 1, Arabidopsis thaliana|Rep: Peptidyl-prolyl cis-trans isomerase - Arabidopsis thaliana (Mouse-ear cress), partial (32%) DT083826 Glycine _(—) soja _(—) 260886 261121 NA release_2 CB063628 GMGI.071508 271592 271900 similar to UniRef100_A7PM52 Cluster: Chromosome chr14 scaffold_21 whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: = partial (2%) CB063628 Glycine _(—) max_release_2 271592 271928 NA TA5835_34305 Lotus _(—) japonicus _(—) 273868 275906 Vegetative cell wall protein release_1 gp1-like [Oryza sativa (japonica cultivar-group) TC32024 LJGI.070108 275152 275906 similar to UniRef100_A7PM52 Cluster: Chromosome chr14 scaffold_21, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr14 scaffold_21, whole genome shotgun sequence - Vitis vinifera (Grape), partial (9%) TC252667 GMGI.071508 275739 276506 similar to UniRef100_A7PM52 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (12%) AW311416 Glycine _(—) max_release_2 276269 276455 NA WmFPC_Contig850 99810 475910 NA CV534998 Phaseolus _(—) vulgaris 288050 288585 UniRef100_A7PM50 Chromosome chr14 scaffold_21, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7PM50_VITVI 6.00E−39 TA75806_3847 Glycine _(—) max_release_2 288290 290376 Arabidopsis thaliana genomic DNA = chromosome 3 = P1 clone: MGF10 [Arabidopsis thaliana (Mouse-ear cress)] TC276120 GMGI.071508 288290 290376 similar to UniRef100_A7PM50 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (62%) BI786388 GMGI.071508 291666 292088 similar to UniRef100_A7PM49 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (7%) BI786388 Glycine _(—) max_release_2 291666 292099 NA TA63308_3847 Glycine _(—) max_release_2 291633 294397 NA TC243765 GMGI.071508 293681 294426 weakly similar to UniRef100_Q0JDM0 Cluster: Os04g0394300 protein; n = 1; Oryza sativa Japonica Group|Rep: Os04g0394300 protein - Oryza sativa subsp. japonica (Rice) = partial (3%) TA6412_34305 Lotus _(—) japonicus _(—) 293803 294412 NA release_1 TC24112 LJGI.070108 293803 294412 NA CA899930 Phaseolus _(—) coccineus _(—) 294054 294263 NA release_2 TA3887_3886 Phaseolus _(—) coccineus _(—) 302301 303033 Hypothetical protein release_2 MJH23.3 [Arabidopsis thaliana (Mouse-ear cress)] AW705271 Glycine _(—) max_release_2 302299 303855 Hypothetical protein MJH23.3 [Arabidopsis thaliana (Mouse-ear cress)] TC237313 GMGI.071508 303227 306007 similar to UniRef100_A7PM30 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (54%) TA61594_3847 Glycine _(—) max_release_2 303227 306056 Similarity to RNA binding protein [Arabidopsis thaliana (Mouse-ear cress)] asmbl_11859 Vigna _(—) unguiculata 303952 305921 NA toGm05 DAGchainer 30059 580791 Ks0.2335 BU544029 Glycine _(—) max_release_2 305220 305762 NA TC23280 LJGI.070108 305373 305839 similar to UniRef100_A7PM30 Cluster: Chromosome chr14 scaffold_21, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr14 scaffold_21, whole genome shotgun sequence - Vitis vinifera (Grape), partial (17%) AI461058 Glycine _(—) max_release_2 305614 305834 NA BE555571 Glycine _(—) max_release_2 305656 306011 NA NGMAX008197032 SEQ. LISTING 314847 315148 SEQ ID NO: 52 asmbl_11860 Vigna _(—) unguiculata 319622 320527 NA EV270366 GMGI.071508 319893 320575 similar to UniRef100_P15792 Cluster: Protein kinase PVPK-1; n = 1; Phaseolus vulgaris|Rep: Protein kinase PVPK-1 - Phaseolus vulgaris (Kidney bean) (French bean) = partial (34%) J04555 Phaseolus _(—) vulgaris _(—) 318937 322709 Protein kinase PVPK-1 release_2 [Phaseolus vulgaris (Kidney bean) (French bean)] TA11578_34305 Lotus _(—) japonicus _(—) 320355 322024 Protein kinase PVPK-1 release_1 [Phaseolus vulgaris (Kidney bean) (French bean)] TC35252 LJGI.070108 320355 322381 homologue to UniRef100_P15792 Cluster: Protein kinase PVPK-1, n = 1, Phaseolus vulgaris|Rep: Protein kinase PVPK-1 - Phaseolus vulgaris (Kidney bean) (French bean), partial (48%) Pvcon4227 Phaseolus _(—) vulgaris 320098 322709 UniRef100_P15792 Protein kinase PVPK-1 n = 1 Tax = Phaseolus vulgaris RepID = KPK1_PHAVU E−0 CA900819 Phaseolus _(—) coccineus _(—) 325129 325547 Sucrase-like protein release_2 [Arabidopsis thaliana (Mouse-ear cress)] CA900820 Phaseolus _(—) coccineus _(—) 325119 328122 AT3g27570/MMJ24_12 release_2 [Arabidopsis thaliana (Mouse-ear cress)] TC269193 GMGI.071508 325136 329359 weakly similar to UniRef100_A7PM27 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (47%) TA4354_3885 Phaseolus _(—) vulgaris _(—) 325476 329154 AT5g40510/MNF13_30 release_2 [Arabidopsis thaliana (Mouse-ear cress)] asmbl_11861 Vigna _(—) unguiculata 326881 329154 NA CF920945 Glycine _(—) max_release_2 326967 329359 AT3g27570/MMJ24_12 [Arabidopsis thaliana (Mouse-ear cress)] SATT723 337605 337828 Satt723 ePCR 337605 337828 Map3.0 SSR L/Gm19 cM: 1.5 TC244213 GMGI.071508 354373 354996 similar to UniRef100_A7PL06 Cluster: Chromosome chr7 scaffold_20 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr7 scaffold_20 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (17%) BU090380 Glycine _(—) max_release_2 354683 354871 NA BP058294 Lotus _(—) japonicus _(—) 355950 356319 Protein ycf2 [Lotus release_1 japonicus] Pvcon2444 Phaseolus _(—) vulgaris 354593 360732 UniRef100_A7PL07 Chromosome chr7 scaffold_20, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7PL07_VITVI 1.00E−144 asmbl_11862 Vigna _(—) unguiculata 359273 359896 NA CA800649 Glycine _(—) max_release_2 377994 379933 AT3g01590/F4P13_13 [Arabidopsis thaliana (Mouse-ear cress)] TC245493 GMGI.071508 377994 381638 similar to UniRef100_A7PM21 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (96%) CO984617 Glycine _(—) max_release_2 379899 381537 At5g14500 [Arabidopsis thaliana (Mouse-ear cress)] M0114388 SEQ. LISTING 381308 380486 SEQ ID NO: 8 AW704585 Glycine _(—) max_release_2 381210 381673 At5g14500 [Arabidopsis thaliana (Mouse-ear cress)] TC248588 GMGI.071508 383419 383857 NA asmbl_11863 Vigna _(—) unguiculata 383428 384088 NA TC126554 MTGI.071708 383593 384668 weakly similar to UniRef100_Q940C3 Cluster: AT3g27530/MMJ24_7, n = 2, Arabidopsis thaliana|Rep: AT3g27530/MMJ24_7 - Arabidopsis thaliana (Mouse-ear cress), partial (38%) AJ002216 Pisum _(—) sativum _(—) 384088 384751 Emb|CAA07228.1 release_2 [Arabidopsis thaliana (Mouse-ear cress)] BI702257 GMGI.071508 384067 384789 similar to UniRef100_Q940C3 Cluster: AT3g27530/MMJ24_7; n = 2; Arabidopsis thaliana|Rep: AT3g27530/MMJ24_7 - Arabidopsis thaliana (Mouse-ear cress) = partial (14%) BG451913 MTGI.071708 386353 388007 similar to UniRef100_Q9LT59 Cluster: Emb|CAA07228.1, n = 1, Arabidopsis thaliana|Rep: Emb|CAA07228.1 - Arabidopsis thaliana (Mouse-ear cress), partial (19%) CV533025 Phaseolus _(—) vulgaris 388647 389345 UniRef100_UPI000016357E GC6 (GOLGIN CANDIDATE 6) binding/ protein transporter Tax = n = 1 RepID = UPI000016357E 6.00E−27 AV777312 LJGI.070108 389152 391279 similar to UniRef100_Q9LT59 Cluster: Emb|CAA07228.1, n = 1, Arabidopsis thaliana|Rep: Emb|CAA07228.1 - Arabidopsis thaliana (Mouse-ear cress), partial (19%) BM187543 GMGI.071508 394984 395407 similar to UniRef100_A7PM13 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (36%) BM187543 Glycine _(—) max_release_2 394984 395559 Gb|AAF01546.1 [Arabidopsis thaliana [Mouse-ear cress)] DN652256 LJGI.070108 395487 395708 similar to UniRef100_A7P4B1 Cluster: Chromosome chr1 scaffold_5, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr1 scaffold_5, whole genome shotgun sequence - Vitis vinifera (Grape), partial (19%) DT044393 Arachis _(—) hypogaea _(—) 395462 395746 Cluster: Hypothetical release_5 protein T23K23.27, n = 1, Arabidopsis thaliana|Rep: Hypothetical protein T23K23.27 - Arabidopsis thaliana (Mouse-ear cress) FD789910 Phaseolus _(—) vulgaris 395555 395927 UniRef100_A7P4B1 Chromosome chr1 scaffold_5, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7P4B1_VITVI 2.00E−59 EH259382 GMGI.071508 395577 396156 similar to UniRef100_A7PM13 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (34%) TA69305_3847 Glycine _(—) max_release_2 403237 404175 NA TC243910 GMGI.071508 403237 404175 similar to UniRef100_A7PM14 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (5%) CA785084 Glycine _(—) soja _(—) 403526 404055 NA release_2 CV541170 Phaseolus _(—) vulgaris _(—) 404688 406556 UniRef100_Q9LT57 Emb|CAB45506.1 n = 1 Tax = Arabidopsis thaliana RepID = Q9LT57_ARATH 1.00E−113 BF071095 GMGI.071508 406510 407127 similar to UniRef100_Q9LT57 Cluster: Emb|CAB45506.1; n = 1; Arabidopsis thaliana|Rep: Emb|CAB45506.1 - Arabidopsis thaliana (Mouse-ear cress) = partial (8%) BF071095 Glycine _(—) max_release_2 406527 407127 NA BM270669 Glycine _(—) max_release_2 409910 410532 NA BM270669 GMGI.071508 410045 410532 similar to UniRef100_A7PM16 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (9%) BG550673 GMGI.071508 421541 422250 similar to UniRef100_A7PM12 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (26%) BG550673 Glycine _(—) max_release_2 421541 422354 Hypothetical protein F18O22_260 [Arabidopsis thaliana (Mouse-ear cress)] BU551363 Glycine _(—) max_release_2 422150 422745 SEQ ID NO: 9 CD407423 Glycine _(—) max_release_2 423719 423842 NA M0205350 SEQ Listing 424095 423776 SEQ ID NO: 10 EV270239 GMGI.071508 425649 426181 similar to UniRef100_Q0WVR7 Cluster: TRNA synthase- like protein; n = 1; Arabidopsis thaliana|Rep: TRNA synthase-like protein - Arabidopsis thaliana (Mouse-ear cress) = partial (5%) BI424448 GMGI.071508 451332 451679 similar to UniRef100_P82353 Cluster: Non-specific lipid- transfer protein 2; n = 1; Prunus armeniaca|Rep: Non-specific lipid-transfer protein 2 - Prunus armeniaca (Apricot) = partial (68%) TA49179_3847 Glycine _(—) max_release_2 451332 451827 Nonspecific lipid-transfer protein 2 [Prunus armeniaca (Apricot)] TC252453 GMGI.071508 451397 451828 weakly similar to UniRef100_Q43681 Cluster: Probable non- specific lipid-transfer protein AKCS9 precursor; n = 1; Vigna unguiculata|Rep: Probable non-specific lipid-transfer protein AKCS9 precursor - Vigna unguiculata (Cowpea) = partial (86%) BE609938 Glycine _(—) max_release_2 451607 451756 Probable lipid transfer protein family protein [Tamarix androssowii] BQ612382 Glycine _(—) max_release_2 451777 452217 NA M0102027 466228 466889 SEQ ID NO: 11 Pvcon7917 Phaseolus _(—) vulgaris 466120 467338 UniRef100_A5C9E2 Putative uncharacterized protein n = 1 Tax = Vitis vinifera RepID = A5C9E2_VITVI 6.00E−44 asmbl_11864 Vigna _(—) unguiculata 467520 468191 NA TA49596_3847 Glycine _(—) max_release_2 470086 472059 Methionine aminopeptidase 2B [Arabidopsis thaliana (Mouse-ear cress)] TC255857 GMGI.071508 470086 476828 homologue to UniRef100_A7PXX3 Cluster: Methionine aminopeptidase; n = 1; Vitis vinifera|Rep: Methionine aminopeptidase - Vitis vinifera (Grape) = partial (91%) FD792539 Phaseolus _(—) vulgaris 472774 475674 UniRef100_A7PXX3 Methionine aminopeptidase n = 1 Tax = Vitis vinifera RepID = A7PXX3_VITVI 5.00E−56 TA3829_3848 Glycine _(—) soja _(—) 471918 476623 Methionine aminopeptidase release_2 2B [Arabidopsis thaliana (Mouse-ear cress)] BU765955 Glycine _(—) max_release_2 472787 475846 Methionine aminopeptidase 2B [Arabidopsis thaliana (Mouse-ear cress)]. SEQ ID NO: 12 EG530516 Arachis _(—) hypogaea _(—) 472835 476690 Cluster: Methionine release_5 aminopeptidase 2B, n = 1, Arabidopsis thaliana|Rep: Methionine aminopeptidase 2B - Arabidopsis thaliana (Mouse-ear cress) AV425234 LJGI.070108 475562 475924 homologue to UniRef100_A7PXX3 Cluster: Methionine aminopeptidase, n = 1, Vitis vinifera|Rep: Methionine aminopeptidase - Vitis vinifera (Grape), partial (22%) TA49598_3847 Glycine _(—) max_release_2 474794 476709 Methionine aminopeptidase 2B [Arabidopsis thaliana (Mouse-ear cress)] FD797260 Phaseolus _(—) vulgaris 475768 476654 UniRef100_A7PXX3 Methionine aminopeptidase n = 1 Tax = Vitis vinifera RepID = A7PXX3_VITVI 6.00E−55 BE823844 Glycine _(—) max_release_2 475751 476828 Methionine aminopeptidase 2B [Arabidopsis thaliana (Mouse-ear cress)] BG726070 Glycine _(—) max_release_2 476668 476807 NA BQ080926 GMGI.071508 480002 480636 similar to UniRef100_A7PY54 Cluster: Chromosome chr15 scaffold_37 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr15 scaffold_37 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (39%) TA69442_3847 Glycine _(—) max_release_2 480002 481069 Hypothetical protein F22I13.40 [Arabidopsis thaliana (Mouse-ear cress)] TC262427 GMGI.071508 480002 481069 similar to UniRef100_A7P8Q6 Cluster: Chromosome chr3 scaffold_8 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr3 scaffold_8 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (20%) BU548976 Glycine _(—) max_release_2 481474 481970 Multi antimicrobial extrusion protein MatE [Medicago truncatula (Barrel medic)] CX547082 Glycine _(—) max_release_2 481345 482173 Multi antimicrobial extrusion protein MatE [Medicago truncatula (Barrel medic)] TC236122 GMGI.071508 481300 482612 NA TA57759_3847 Glycine _(—) max_release_2 481300 482627 Multi antimicrobial extrusion protein MatE [Medicago truncatula (Barrel medic)] AV420909 LJGI.070108 481846 482201 weakly similar to UniRef100_A7QTE8 Cluster: Chromosome undetermined scaffold_167, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome undetermined scaffold_167, whole genome shotgun sequence - Vitis vinifera (Grape), partial (24%) AW597322 Glycine _(—) max_release_2 481965 482825 Multi antimicrobial extrusion protein MatE [Medicago truncatula (Barrel medic)] BM270610 Glycine _(—) max_release_2 482034 483008 Multi antimicrobial extrusion protein MatE [Medicago truncatula (Barrel medic)] BI972603 GMGI.071508 482632 483190 weakly similar to UniRef100_A7P3G6 Cluster: Chromosome chr1 scaffold_5 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr1 scaffold_5 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (20%) BI972603 Glycine _(—) max_release_2 482632 484113 Multi antimicrobial extrusion protein MatE [Medicago truncatula (Barrel medic)] TA66198_3847 Glycine _(—) max_release_2 482595 484230 Multi antimicrobial extrusion protein MatE [Medicago truncatula (Barrel medic)] TC253566 GMGI.071508 482648 484405 weakly similar to UniRef100_A7QTE8 Cluster: Chromosome undetermined scaffold_167 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome undetermined scaffold_167 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (44%) asmbl_11865 Vigna _(—) unguiculata 482937 484289 NA BG881371 Glycine _(—) max_release_2 483075 484230 Multi antimicrobial extrusion protein MatE [Medicago truncatula (Barrel medic)] WmFPC_Contig7443 384071 598745 NA AW695419 MTGI.071708 491367 494466 similar to UniRef100_A7PU69 Cluster: Chromosome chr7 scaffold_31, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr7 scaffold_31, whole genome shotgun sequence - Vitis vinifera (Grape), partial (11%) BF645755 MTGI.071708 494870 497474 similar to UniRef100_A7PU69 Cluster: Chromosome chr7 scaffold_31, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr7 scaffold_31, whole genome shotgun sequence - Vitis vinifera (Grape), partial (14%) BE475242 GMGI.071508 497000 497327 similar to UniRef100_A7NWE7 Cluster: Chromosome chr5 scaffold_2 whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: = partial (1%) BE475242 Glycine _(—) max_release_2 497000 497549 Hypothetical protein At3g23590/MDB19_8 [Arabidopsis thaliana (Mouse-ear cress)] BW611072 LJGI.070108 497387 497795 similar to UniRef100_A7PU69 Cluster: Chromosome chr7 scaffold_31, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr7 scaffold_31, whole genome shotgun sequence - Vitis vinifera (Grape), partial (10%) BQ613050 Glycine _(—) max_release_2 497409 498014 ORF protein [Arabidopsis thaliana (Mouse-ear cress)] CV541244 Phaseolus _(—) vulgaris 500143 500464 UniRef100_A9PGX2 Putative uncharacterized protein n = 1 Tax = Populus trichocarpa RepID = A9PGX2_POPTR 3.00E−28 CX856527 Glycine _(—) max_release_2 501517 501735 NA BG839076 Glycine _(—) max_release_2 503126 505209 F2P3.12 protein [Arabidopsis thaliana (Mouse-ear cress)] FD790090 Phaseolus _(—) vulgaris 503370 505191 No significant hit (e−20) TC236383 GMGI.071508 503107 505675 similar to UniRef100_O82505 Cluster: Elongation factor Ts; n = 1; Arabidopsis thaliana|Rep: Elongation factor Ts - Arabidopsis thaliana (Mouse-ear cress) = partial (32%) TA56246_3847 Glycine _(—) max_release_2 503107 505848 Ethylene-responsive elongation factor EF-Ts precursor [Lycopersicon esculentum (Tomato)] TC239475 GMGI.071508 503126 506560 similar to UniRef100_Q9SWW0 Cluster: Ethylene- responsive elongation factor EF-Ts precursor; n = 1; Solanum lycopersicum|Rep: Ethylene-responsive elongation factor EF-Ts precursor Solanum lycopersicum (Tomato) (Lycopersicon esculentum) = partial (74%) TA56245_3847 Glycine _(—) max_release_2 505512 506546 Ethylene-responsive elongation factor EF-Ts precursor [Lycopersicon esculentum (Tomato)] BG839060 Glycine _(—) max_release_2 505661 506530 At4g11120 [Arabidopsis thaliana (Mouse-ear cress)] CV543527 Phaseolus _(—) vulgaris _(—) 508539 508771 Eukaryotic translation release_2 initiation factor 5 [Phaseolus vulgaris (Kidney bean) (French bean)] CD393454 Glycine _(—) max_release_2 510651 511000 Ribosomal protein L22 [Glycine max (Soybean)] TC245517 GMGI.071508 510651 511270 homologue to UniRef100_O48879 Cluster: Ribosomal protein L22; n = 1; Glycine max|Rep: Ribosomal protein L22 - Glycine max (Soybean) = partial (80%) asmbl_11866 Vigna _(—) unguiculata 510868 511269 NA TA51206_3847 Glycine _(—) max_release_2 510702 512712 Ribosomal protein L22 [Glycine max (Soybean)] TC249077 GMGI.071508 510771 512771 homologue to UniRef100_O48879 Cluster: Ribosomal protein L22; n = 1; Glycine max|Rep: Ribosomal protein L22 - Glycine max (Soybean) = partial (98%) BG316244 Glycine _(—) max_release_2 511015 512722 Ribosomal protein L22 [Glycine max (Soybean)] BQ155270 MTGI.071708 513084 514936 similar to UniRef100_A7PR59 Cluster: Chromosome chr14 scaffold_26, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr14 scaffold_26, whole genome shotgun sequence - Vitis vinifera (Grape), partial (52%) TC30151 LJGI.070108 514647 516395 similar to UniRef100_A7PR59 Cluster: Chromosome chr14 scaffold_26, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr14 scaffold_26, whole genome shotgun sequence - Vitis vinifera (Grape), partial (29%) BP044357 Lotus _(—) japonicus _(—) 514647 516409 S-locus protein 8 [Brassica release_1 campestris (Field mustard)] CB540591 Phaseolus _(—) vulgaris 514839 516355 No significant hit (e−20) TA65114_3847 Glycine _(—) max_release_2 523413 524053 At1g22990/F19G10_22 [Arabidopsis thaliana (Mouse-ear cress)] TC259745 GMGI.071508 523413 524067 similar to UniRef100_A7P3I8 Cluster: Chromosome chr1 scaffold_5 = whole genome shotgun sequence; n = 2; Vitis vinifera|Rep: Chromosome chr1 scaffold_5 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (56%) TA4332_47247 Lotus _(—) corniculatus _(—) 529321 530051 Actin-11 related cluster release_1 TA6031_34305 Lotus _(—) japonicus _(—) 529321 530051 Actin [Striga asiatica] release_1 TC32457 LJGI.070108 529321 530051 homologue to UniRef100_P30167 Cluster: Actin-58, n = 1, Solanum tuberosum|Rep: Actin-58 - Solanum tuberosum (Potato), partial (39%) AW351005 Glycine _(—) max_release_2 529380 530095 Actin [Striga asiatica] TA43521_3847 Glycine _(—) max_release_2 529306 530175 Actin-11 [Arabidopsis thaliana (Mouse-ear cress)] asmbl_11867 Vigna _(—) unguiculata 529342 530189 NA AU240079 LJGI.070108 529747 530013 homologue to UniRef100_P93372 Cluster: Actin-66, n = 1, Nicotiana tabacum|Rep: Actin-66 - Nicotiana tabacum (Common tobacco), partial (25%) AU240079 Lotus _(—) japonicus _(—) 529747 530039 Actin-11 [Arabidopsis release_1 thaliana (Mouse-ear cress)] EE127018 Arachis _(—) hypogaea _(—) 529933 530285 Cluster: Hypothetical release_5 protein, n = 1, Oryza sativa (indica cultivar-group)|Rep: Hypothetical protein - Oryza sativa subsp. indica (Rice) TC240040 GMGI.071508 529306 531078 homologue to UniRef100_P02581 Cluster: Actin-1; n = 1; Glycine max|Rep: Actin-1 - Glycine max (Soybean) = complete AW666288 Glycine _(—) max_release_2 529980 530789 Actin [Phaseolus acutifolius (Tepary bean)] TA43509_3847 Glycine _(—) max_release_2 529888 530911 Actin [Glycine max (Soybean)] TA6074_34305 Lotus _(—) japonicus _(—) 530031 531095 Actin-1 [Sorghum bicolor release_1 (Sorghum) (Sorghum vulgare)] TC26188 LJGI.070108 530031 531095 homologue to UniRef100_A1Y2A0 Cluster: Actin, n = 1, Aegiceras corniculatum|Rep: Actin - Aegiceras corniculatum, partial (81%) BM142797 Glycine _(—) max_release_2 530212 531095 Actin [Trifolium pratense (Red clover)] BP036880 Lotus _(—) japonicus _(—) 530235 531095 Actin/actin-like [Medicago release_1 truncatula (Barrel medic)] AW349632 Glycine _(—) max_release_2 533113 533701 NA AI900119 Glycine _(—) max_release_2 533044 534995 NA TA51800_3847 Glycine _(—) max_release_2 533054 535063 NA TC241826 GMGI.071508 533055 535063 similar to UniRef100_Q2Z1Y5 Cluster: Pm52 protein; n = 1; Prunus mume|Rep: Pm52 protein - Prunus mume (Japanese flowering apricot) = partial (73%) BU494245 LJGI.070108 533191 534994 weakly similar to UniRef100_Q2Z1Y5 Cluster: Pm52 protein, n = 1, Prunus mume|Rep: Pm52 protein - Prunus mume (Japanese flowering apricot), partial (59%) AI440735 Glycine _(—) max_release_2 534517 535020 NA AI440735 GMGI.071508 534522 535020 similar to UniRef100_Q2Z1Y5 Cluster: Pm52 protein; n = 1; Prunus mume|Rep: Pm52 protein - Prunus mume (Japanese flowering apricot) = partial (41%) TC250013 GMGI.071508 536842 537680 UniRef100_Q8L7J4 Cluster: Pyruvate kinase; n = 1; Glycine max|Rep: Pyruvate kinase - Glycine max (Soybean) = partial (29%) TA10574_34305 Lotus _(—) japonicus _(—) 537149 537628 Pyruvate kinase [Glycine release_1 max (Soybean)] TC26632 LJGI.070108 537149 537628 homologue to UniRef100_Q42806 Cluster: Pyruvate kinase, cytosolic isozyme, n = 1, Glycine max|Rep: Pyruvate kinase, cytosolic isozyme - Glycine max (Soybean), partial (26%) CV536725 Phaseolus _(—) vulgaris _(—) 537147 537846 Pyruvate kinase = cytosolic release_2 isozyme [Glycine max (Soybean)] asmbl_11868 Vigna _(—) unguiculata 537127 538325 NA TC25282 LJGI.070108 537149 538489 homologue to UniRef100_Q8L7J4 Cluster: Pyruvate kinase, n = 1, Glycine max|Rep: Pyruvate kinase - Glycine max (Soybean), partial (29%) TA47094_3847 Glycine _(—) max_release_2 536842 539314 Pyruvate kinase [Glycine max (Soybean)] Pvcon4373 Phaseolus _(—) vulgaris 537147 539113 UniRef100_Q42806 Pyruvate kinase, cytosolic isozyme n = 1 Tax = Glycine max RepID = KPYC_SOYBN E−0 TC124922 MTGI.071708 537491 538783 homologue to UniRef100_Q42806 Cluster: Pyruvate kinase, cytosolic isozyme, n = 1, Glycine max|Rep: Pyruvate kinase, cytosolic isozyme - Glycine max (Soybean), partial (64%) BF598352 Glycine _(—) soja _(—) 538308 538971 Pyruvate kinase [Citrus release_2 sinensis (Sweet orange)] BG044770 Glycine _(—) soja _(—) 538624 539149 Pyruvate kinase [Citrus release_2 sinensis (Sweet orange)] TC249941 GMGI.071508 538549 539314 UniRef100_Q8L7J4 Cluster: Pyruvate kinase; n = 1; Glycine max|Rep: Pyruvate kinase - Glycine max (Soybean) = partial (37%) BE608312 Glycine _(—) max_release_2 542536 544875 Hypothetical protein [Arabidopsis thaliana (Mouse-ear cress)] TC253996 GMGI.071508 542045 546856 similar to UniRef100_A7QNQ5 Cluster: Chromosome undetermined scaffold_133 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome undetermined scaffold_133 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (80%) TC258772 GMGI.071508 548268 548805 NA CV533614 Phaseolus _(—) vulgaris 548540 548638 No significant hit TA57756_3847 Glycine _(—) max_release_2 548268 551375 Putative microtubule- severing protein subunit [Oryza sativa (japonica cultivar-group)] TC239891 GMGI.071508 548323 551375 similar to UniRef100_A7QNQ6 Cluster: Chromosome undetermined scaffold_133 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome undetermined scaffold_133 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (12%) EH221990 GMGI.071508 550796 551633 weakly similar to UniRef100_A7QNQ6 Cluster: Chromosome undetermined scaffold_133 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome undetermined scaffold_133 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (7%) EV263369 GMGI.071508 552842 553615 similar to UniRef100_A8D2Q2 Cluster: ATP synthase protein 8; n = 1; Caranx ignobilis|Rep: ATP synthase protein 8 - Caranx ignobilis = partial (37%) BU964969 Glycine _(—) max_release_2 556336 556943 NA BU964969 GMGI.071508 556494 556943 similar to UniRef100_Q9MYM4 Cluster: Lysosomal alpha- glucosidase precursor; n = 1; Bos taurus|Rep: Lysosomal alpha-glucosidase = partial (1%) EH221989 GMGI.071508 562783 563692 homologue to UniRef100_A7QNQ6 Cluster: Chromosome undetermined scaffold_133 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome undetermined scaffold_133 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (3%) AW831441 GMGI.071508 573069 573567 NA AW831441 Glycine _(—) max_release_2 573069 573639 NA TA6761_34305 Lotus _(—) japonicus _(—) 573706 580487 Sphingosine kinase [Lotus release_1 japonicus] TC20288 LJGI.070108 573706 580487 UniRef100_Q5KR50 Cluster: Sphingosine kinase, n = 1, Lotus japonicus|Rep: Sphingosine kinase - Lotus japonicus, complete TC122322 MTGI.071708 574490 580620 homologue to UniRef100_Q5KR50 Cluster: Sphingosine kinase, n = 1, Lotus japonicus|Rep: Sphingosine kinase - Lotus japonicus, partial (66%) BI701010 Glycine _(—) max_release_2 577145 579375 Sphingosine kinase [Lotus japonicus] Pvcon3123 Phaseolus _(—) vulgaris 577107 580468 UniRef100_Q5KR50 Sphingosine kinase n = 1 Tax = Lotus japonicus RepID = Q5KR50_LOTJA E−0 TA49258_3847 Glycine _(—) max_release_2 579511 580791 Sphingosine kinase [Lotus japonicus] TC235674 GMGI.071508 579511 580791 homologue to UniRef100_Q5KR50 Cluster: Sphingosine kinase; n = 1; Lotus japonicus|Rep: Sphingosine kinase - Lotus japonicus = partial (26%) BI969866 Glycine _(—) max_release_2 579600 580756 Sphingosine kinase [Lotus japonicus] EH043869 Arachis _(—) stenosperma _(—) 579729 580660 Cluster: Sphingosine release_5 kinase, n = 1, Lotus japonicus|Rep: Sphingosine kinase - Lotus japonicus BQ786742 Glycine _(—) max_release_2 580594 580719 NA BM108235 Glycine _(—) max_release_2 581688 582006 NA AW508189 Glycine _(—) max_release_2 581725 582244 Hypothetical protein [Arabidopsis thaliana (Mouse-ear cress)] TC238711 GMGI.071508 581688 582562 similar to UniRef100_A7QNQ7 Cluster: Chromosome undetermined scaffold_133 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome undetermined scaffold_133 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (50%) TA46155_3847 Glycine _(—) max_release_2 581745 582556 Hypothetical protein [Arabidopsis thaliana (Mouse-ear cress)] AW278369 GMGI.071508 581988 582389 similar to UniRef100_A7QNQ7 Cluster: Chromosome undetermined scaffold_133 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome undetermined scaffold_133 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (44%) AW278369 Glycine _(—) max_release_2 581988 582418 Hypothetical protein [Arabidopsis thaliana (Mouse-ear cress)] CD394810 Glycine _(—) max_release_2 582134 582328 NA BG047332 Glycine _(—) max_release_2 591288 592013 OSJNBb0065L13.3 protein [Oryza sativa (japonica cultivar-group)] TC272805 GMGI.071508 591358 592013 similar to UniRef100_A7NXM8 Cluster: Chromosome chr5 scaffold_2 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr5 scaffold_2 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (15%) BW599171 LJGI.070108 593399 593875 weakly similar to UniRef100_A7PT63 Cluster: Chromosome chr8 scaffold_29, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr8 scaffold_29, whole genome shotgun sequence - Vitis vinifera (Grape), partial (24%) BE057829 Glycine _(—) max_release_2 606858 607008 NA TC275159 GMGI.071508 606858 607456 NA BE612118 GMGI.071508 615853 616253 weakly similar to UniRef100_A7GPV4 Cluster: Citrate transporter; n = 1; Bacillus cereus subsp. cytotoxis NVH 391-98|Rep: Citrate transporter - Bacillus cereus subsp. cytotoxis (strain NVH 391- 98) = partial (5%) BE612118 Glycine _(—) max_release_2 615869 616269 NA CA910895 Phaseolus _(—) coccineus _(—) 622174 622531 Arabidopsis thaliana release_2 genomic DNA, chromosome 5, P1 clone: MPO12 [Arabidopsis thaliana (Mouse-ear cress)] BU763992 Glycine _(—) max_release_2 625192 625591 NA TA51978_3847 Glycine _(—) max_release_2 625330 626304 Putative ethylene- responsive protein [Oryza sativa (japonica cultivar- group)] TC236117 GMGI.071508 625330 626304 similar to UniRef100_A7PM86 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (43%) TC263881 GMGI.071508 625192 627651 similar to UniRef100_A7PM86 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (76%) TA51979_3847 Glycine _(—) max_release_2 625252 627642 Putative ethylene response protein [Capsicum chinense (Scotch bonnet) (Bonnet pepper)] TC236300 GMGI.071508 625318 627642 similar to UniRef100_A7PM86 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (98%) CA910548 Phaseolus _(—) coccineus _(—) 625559 627607 Putative ethylene response release_2 protein [Capsicum chinense (Scotch bonnet) (Bonnet pepper)] Pvcon5808 Phaseolus _(—) vulgaris 625567 627610 UniRef100_A7PM86 Chromosome chr14 scaffold_21, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7PM86_VITVI 2.00E−77 EV269595 GMGI.071508 627204 627569 similar to UniRef100_A7PM86 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (29%) BI273677 Glycine _(—) max_release_2 637550 637816 NA BP049107 Lotus _(—) corniculatus _(—) 647584 649419 Cinnamoyl CoA reductase- release_1 like protein related cluster TC258382 GMGI.071508 646415 652371 weakly similar to UniRef100_A7PM88 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (72%) TA50222_3847 Glycine _(—) max_release_2 646722 652222 Cinnamoyl CoA reductase- like protein [Arabidopsis thaliana (Mouse-ear cress)] SATT495 650288 650531 Satt495 ePCR 650288 650531 Map3.0 SSR L/Gm19 cM: 2.7 AW099618 GMGI.071508 649276 652222 weakly similar to UniRef100_A7PM88 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (23%) TA50296_3847 Glycine _(—) max_release_2 674409 676421 NA BQ629031 Glycine _(—) max_release_2 674669 676494 NA BM520842 Glycine _(—) soja _(—) 674685 676538 NA release_2 TC264557 GMGI.071508 674741 676494 NA BU765059 Glycine _(—) max_release_2 674828 676698 NA BU765059 GMGI.071508 674925 676698 weakly similar to UniRef100_A7L4B0 Cluster: Protein kinase; n = 1; Carica papaya|Rep: Protein kinase Carica papaya (Papaya) = partial (6%) TC264815 GMGI.071508 674409 678111 weakly similar to UniRef100_A7L4B0 Cluster: Protein kinase; n = 1; Carica papaya|Rep: Protein kinase - Carica papaya (Papaya) = partial (14%) asmbl_11869 Vigna _(—) unguiculata 676473 676672 NA TA50295_3847 Glycine _(—) max_release_2 674775 678957 NA Pvcon1987 Phaseolus _(—) vulgaris 674506 679702 UniRef100_A7L4B0 Protein kinase n = 1 Tax = Carica papaya RepID = A7L4B0_CARPA 1.00E−127 BM528477 Glycine _(—) max_release_2 676507 678111 NA TA11531_47247 Lotus _(—) corniculatus _(—) 676692 678714 Protein kinase-like protein release_1 related cluster TA13031_34305 Lotus _(—) japonicus _(—) 676692 678714 Hypothetical protein release_1 At5g14720 [Arabidopsis thaliana (Mouse-ear cress)] TC31122 LJGI.070108 676701 678714 similar to UniRef100_A7L4B0 Cluster: Protein kinase, n = 1, Carica papaya|Rep: Protein kinase - Carica papaya (Papaya), partial (14%) TC255388 GMGI.071508 679127 681361 homologue to UniRef100_A7PM90 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (44%) TC124284 MTGI.071708 679117 681419 homologue to UniRef100_A7PM90 Cluster: Chromosome chr14 scaffold_21, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr14 scaffold_21, whole genome shotgun sequence - Vitis vinifera (Grape), partial (48%) DV565290 Phaseolus _(—) vulgaris 681368 681460 No significant hit (e−20) toGm05 DAGchainer 603011 803108 Ks0.2166 NP7265365 MTGI.071708 703588 713159 GB|AC124951.19|ABE84834.1 ATPase, E1-E2 type, Peptidase M, neutral zinc metallopeptidases, zinc- binding site BF325038 Glycine _(—) max_release_2 711165 712911 ATPase = E1-E2 type; Peptidase M = neutral zinc metallopeptidases = zinc- binding site [Medicago truncatula (Barrel medic)] FE897117 Phaseolus _(—) vulgaris 715539 715874 UniRef100_Q93VL6 NBS- LRR resistance-like protein J78 n = 1 Tax = Phaseolus vulgaris RepID = Q93VL6_PHAVU 2.00E−47 TC264844 GMGI.071508 731939 732440 weakly similar to UniRef100_A7PD05 Cluster: Chromosome chr17 scaffold_12 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr17 scaffold_12 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (13%) TA67235_3847 Glycine _(—) max_release_2 731939 733078 NA CD404253 GMGI.071508 732439 733078 homologue to UniRef100_A7PM92 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (8%) BU091162 GMGI.071508 737876 738292 NA BU091162 Glycine _(—) max_release_2 737876 738363 NA asmbl_11870 Vigna _(—) unguiculata 740144 741401 NA BI470779 GMGI.071508 740189 741746 similar to UniRef100_Q9XQB0 Cluster: Carbonic anhydrase; n = 1; Vigna radiata var. radiata|Rep: Carbonic anhydrase - Phaseolus aureus (Mung bean) (Vigna radiata) = partial (30%) TA43150_3847 Glycine _(—) max_release_2 740126 742524 Carbonic anhydrase [Phaseolus aureus (Mung bean) (Vigna radiata)] BG509786 GMGI.071508 740265 742434 homologue to UniRef100_Q9XQB0 Cluster: Carbonic anhydrase; n = 1; Vigna radiata var. radiata|Rep: Carbonic anhydrase - Phaseolus aureus (Mung bean) (Vigna radiata) = partial (34%) BG509786 Glycine _(—) max_release_2 740265 742656 Carbonic anhydrase [Zea mays (Maize)] DT083317 Glycine _(—) soja _(—) 740299 742670 Carbonic anhydrase [Zea release_2 mays (Maize)] AW781596 Glycine _(—) max_release_2 740182 742860 Carbonic anhydrase [Phaseolus aureus (Mung bean) (Vigna radiata)] BU089680 Glycine _(—) max_release_2 741070 742671 Carbonic anhydrase [Zea mays (Maize)] BM887226 Glycine _(—) max_release_2 741037 742852 Carbonic anhydrase [Zea mays (Maize)] BU089600 Glycine _(—) max_release_2 741070 742891 Carbonic anhydrase [Zea mays (Maize)] TC23104 LJGI.070108 740127 744319 similar to UniRef100_Q9XQB0 Cluster: Carbonic anhydrase, n = 1, Vigna radiata var. radiata|Rep: Carbonic anhydrase - Phaseolus aureus (Mung bean) (Vigna radiata), partial (98%) TA2934_3885 Phaseolus _(—) vulgaris _(—) 739932 744687 Carbonic anhydrase [Zea release_2 mays (Maize)] TC238511 GMGI.071508 740118 744639 homologue to UniRef100_Q9XQB0 Cluster: Carbonic anhydrase; n = 1; Vigna radiata var. radiata|Rep: Carbonic anhydrase - Phaseolus aureus (Mung bean) (Vigna radiata) = complete TA377_34305 Lotus _(—) japonicus _(—) 740127 744704 Carbonic anhydrase [Zea release_1 mays (Maize)] Pvcon229 Phaseolus _(—) vulgaris 740125 744728 UniRef100_Q9XQB0 Carbonic anhydrase n = 1 Tax = Vigna radiata var. radiata RepID = Q9XQB0_PHAAU 1.00E−176 TA2935_3885 Phaseolus _(—) vulgaris _(—) 740178 744687 Carbonic anhydrase [Zea release_2 mays (Maize)] TA2376_3848 Glycine _(—) soja _(—) 740118 744805 Carbonic anhydrase release_2 [Phaseolus aureus (Mung bean) (Vigna radiata)] TA43157_3847 Glycine _(—) max_release_2 740117 744844 Carbonic anhydrase [Zea mays (Maize)] TA43160_3847 Glycine _(—) max_release_2 741051 744186 Carbonic anhydrase = chloroplast precursor (EC 4.2.1.1) (Carbonate dehydratase) [Contains: Carbonic anhydrase = 27 kDa isoform; Carbonic anhydrase = 25 kDa isoform] [Pisum sativum (Garden pea)] TC135779 MTGI.071708 741364 744530 homologue to UniRef100_P17067 Cluster: Carbonic anhydrase, chloroplast precursor (Carbonate dehydratase) [Contains: Carbonic anhydrase, 27 kDa isoform, Carbonic anhydrase, 25 kDa isoform], n = 1, Pisum sativum|Rep: Carbonic anhydrase, chloroplast precursor (Carbonate dehydratase) [Contains: Carbonic anhydrase, 27 kDa isoform, Carbonic anhydrase, 25 kDa isoform] - Pisum sativum (Garden pea), partial (79%) TA4174_3848 Glycine _(—) soja _(—) 742624 743398 Carbonic anhydrase release_2 [Phaseolus aureus (Mung bean) (Vigna radiata)] Pvcon228 Phaseolus _(—) vulgaris 741374 744687 UniRef100_Q9XQB0 Carbonic anhydrase n = 1 Tax = Vigna radiata var. radiata RepID = Q9XQB0_PHAAU 1.00E−137 TA43163_3847 Glycine _(—) max_release_2 741381 744770 Carbonic anhydrase [Zea mays (Maize)] TC247359 GMGI.071508 741381 744770 homologue to UniRef100_Q9XQB0 Cluster: Carbonic anhydrase; n = 1; Vigna radiata var. radiata|Rep: Carbonic anhydrase - Phaseolus aureus (Mung bean) (Vigna radiata) = partial (62%) BG045644 Glycine _(—) soja _(—) 742643 743622 Carbonic anhydrase = release_2 chloroplast precursor (EC 4.2.1.1) (Carbonate dehydratase) [Contains: Carbonic anhydrase = 27 kDa isoform; Carbonic anhydrase = 25 kDa isoform] [Pisum sativum (Garden pea)] Pvcon227 Phaseolus _(—) vulgaris 741681 744687 UniRef100_Q9XQB0 Carbonic anhydrase n = 1 Tax = Vigna radiata var. radiata RepID = Q9XQB0_PHAAU 1.00E−133 TC124201 MTGI.071708 741922 744665 homologue to UniRef100_P17067 Cluster: Carbonic anhydrase, chloroplast precursor (Carbonate dehydratase) [Contains: Carbonic anhydrase, 27 kDa isoform, Carbonic anhydrase, 25 kDa isoform], n = 1, Pisum sativum|Rep: Carbonic anhydrase, chloroplast precursor (Carbonate dehydratase) [Contains: Carbonic anhydrase, 27 kDa isoform, Carbonic anhydrase, 25 kDa isoform] - Pisum sativum (Garden pea), partial (57%) CB543710 Phaseolus _(—) vulgaris _(—) 742464 744532 Carbonic anhydrase release_2 [Phaseolus aureus (Mung bean) (Vigna radiata)] CB539509 Phaseolus _(—) vulgaris _(—) 742480 744557 Carbonic anhydrase [Zea release_2 mays (Maize)] TC126947 MTGI.071708 742434 744665 homologue to UniRef100_P17067 Cluster: Carbonic anhydrase, chloroplast precursor (Carbonate dehydratase) [Contains: Carbonic anhydrase, 27 kDa isoform, Carbonic anhydrase, 25 kDa isoform], n = 1, Pisum sativum|Rep: Carbonic anhydrase, chloroplast precursor (Carbonate dehydratase) [Contains: Carbonic anhydrase, 27 kDa isoform, Carbonic anhydrase, 25 kDa isoform] - Pisum sativum (Garden pea), partial (51%) asmbl_11871 Vigna _(—) unguiculata 742823 744369 NA asmbl_11872 Vigna _(—) unguiculata 742628 744687 NA asmbl_11874 Vigna _(—) unguiculata 742641 744687 NA TA43165_3847 Glycine _(—) max_release_2 742658 744772 Carbonic anhydrase [Phaseolus aureus (Mung bean) (Vigna radiata)] TC241035 GMGI.071508 742658 744772 homologue to UniRef100_Q9XQB0 Cluster: Carbonic anhydrase; n = 1; Vigna radiata var. radiata|Rep: Carbonic anhydrase - Phaseolus aureus (Mung bean) (Vigna radiata) = partial (38%) TA480_3888 Pisum _(—) sativum _(—) 742823 744641 Carbonic anhydrase, release_2 chloroplast precursor (EC 4.2.1.1) (Carbonate dehydratase) [Contains: Carbonic anhydrase, 27 kDa isoform, Carbonic anhydrase, 25 kDa isoform] [Pisum sativum (Garden pea)] TC240357 GMGI.071508 742650 744828 homologue to UniRef100_Q9XQB0 Cluster: Carbonic anhydrase; n = 1; Vigna radiata var. radiata|Rep: Carbonic anhydrase - Phaseolus aureus (Mung bean) (Vigna radiata) = partial (38%) BE346766 Glycine _(—) max_release_2 743636 744227 Carbonic anhydrase = chloroplast precursor (EC 4.2.1.1) (Carbonate dehydratase) [Contains: Carbonic anhydrase = 27 kDa isoform; Carbonic anhydrase = 25 kDa isoform] [Pisum sativum (Garden pea)] AW596246 Glycine _(—) max_release_2 743636 744243 Carbonic anhydrase [Phaseolus aureus (Mung bean) (Vigna radiata)] BE807206 Glycine _(—) max_release_2 743636 744244 Carbonic anhydrase [Phaseolus aureus (Mung bean) (Vigna radiata)] CB280659 Phaseolus _(—) vulgaris _(—) 743613 744419 Carbonic anhydrase release_2 [Phaseolus aureus (Mung bean) (Vigna radiata)] asmbl_11875 Vigna _(—) unguiculata 743587 744642 NA DT083076 Glycine _(—) soja _(—) 743565 744678 Carbonic anhydrase release_2 [Phaseolus aureus (Mung bean) (Vigna radiata)] TC29040 LJGI.070108 743565 744702 similar to UniRef100_Q9XQB0 Cluster: Carbonic anhydrase, n = 1, Vigna radiata var. radiata|Rep: Carbonic anhydrase - Phaseolus aureus (Mung bean) (Vigna radiata), partial (31%) TA134_47247 Lotus _(—) corniculatus _(—) 743568 744704 Carbonic anhydrase related release_1 cluster TA378_34305 Lotus _(—) japonicus _(—) 743568 744704 Carbonic anhydrase, release_1 prokaryotic and plant [Medicago truncatula (Barrel medic)] TC24201 LJGI.070108 743584 744704 similar to UniRef100_Q9XQB0 Cluster: Carbonic anhydrase, n = 1, Vigna radiata var. radiata|Rep: Carbonic anhydrase - Phaseolus aureus (Mung bean) (Vigna radiata), partial (25%) CB539196 Phaseolus _(—) vulgaris _(—) 743626 744687 Carbonic anhydrase release_2 [Phaseolus aureus (Mung bean) (Vigna radiata)] AV413187 LJGI.070108 744089 744647 similar to UniRef100_P27140 Cluster: Carbonic anhydrase, chloroplast precursor, n = 4, Arabidopsis thaliana|Rep: Carbonic anhydrase, chloroplast precursor - Arabidopsis thaliana (Mouse-ear cress), partial (17%) AV413187 Lotus _(—) japonicus _(—) 744089 744672 Carbonic anhydrase, release_1 chloroplast precursor [Arabidopsis thaliana (Mouse-ear cress)] CD860850 Pisum _(—) sativum _(—) 744145 744641 Carbonic anhydrase, release_2 chloroplast precursor [Arabidopsis thaliana (Mouse-ear cress)] CD403834 Glycine _(—) max_release_2 744076 744732 Carbonic anhydrase = chloroplast precursor [Arabidopsis thaliana (Mouse-ear cress)] CD415400 Glycine _(—) max_release_2 744251 744691 NA asmbl_11873 Vigna _(—) unguiculata 744448 744649 NA CB541850 Phaseolus _(—) vulgaris 747218 747570 No significant hit (e−20) BM953717 Glycine _(—) max_release_2 747199 748912 Peptidase S1 and S6 = chymotrypsin/Hap [Medicago truncatula (Barrel medic)] EH256926 GMGI.071508 747192 749279 homologue to UniRef100_A7Q7E6 Cluster: Chromosome chr18 scaffold_59 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr18 scaffold_59 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (21%) TA51716_3847 Glycine _(—) max_release_2 747191 749327 Putative DegP protease [Oryza sativa (japonica cultivar-group)] TC243148 GMGI.071508 747199 749327 homologue to UniRef100_A7Q7E6 Cluster: Chromosome chr18 scaffold_59 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr18 scaffold_59 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (25%) AV768772 LJGI.070108 747281 749288 homologue to UniRef100_O22609 Cluster: Protease Do-like 1, chloroplast precursor, n = 1, Arabidopsis thaliana|Rep: Protease Do-like 1, chloroplast precursor - Arabidopsis thaliana (Mouse-ear cress), partial (23%) BE807421 Glycine _(—) max_release_2 748776 749688 Peptidase S1 and S6 = chymotrypsin/Hap [Medicago truncatula (Barrel medic)] TA51715_3847 Glycine _(—) max_release_2 747251 752927 Peptidase S1 and S6 = chymotrypsin/Hap [Medicago truncatula (Barrel medic)] TC260884 GMGI.071508 747251 752942 homologue to UniRef100_A7Q7E6 Cluster: Chromosome chr18 scaffold_59 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr18 scaffold_59 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (80%) BE474482 Glycine _(—) max_release_2 751068 752387 Peptidase S1 and S6 = chymotrypsin/Hap [Medicago truncatula (Barrel medic)] BE474482 GMGI.071508 751070 752387 homologue to UniRef100_A7Q7E6 Cluster: Chromosome chr18 scaffold_59 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr18 scaffold_59 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (19%) TC261290 GMGI.071508 755656 757218 similar to UniRef100_A7PM96 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (33%) BG646067 MTGI.071708 756996 759297 similar to UniRef100_A7PM96 Cluster: Chromosome chr14 scaffold_21, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr14 scaffold_21, whole genome shotgun sequence - Vitis vinifera (Grape), partial (33%) BE555567 Glycine _(—) max_release_2 757210 762134 Hypothetical protein [Medicago truncatula (Barrel medic)] BE555567 GMGI.071508 757746 762134 similar to UniRef100_A7PM96 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (31%) BE058948 Glycine _(—) max_release_2 762117 763784 Hypothetical protein [Medicago truncatula (Barrel medic)] BE058948 GMGI.071508 762818 763784 similar to UniRef100_A7PM96 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (25%) TC138874 MTGI.071708 768876 770881 similar to UniRef100_Q40318 Cluster: Coil protein, n = 1, Medicago sativa|Rep: Coil protein - Medicago sativa (Alfalfa), partial (60%) TC124470 MTGI.071708 768770 771318 similar to UniRef100_Q1RU40 Cluster: Lipolytic enzyme, G-D-S-L, n = 1, Medicago truncatula|Rep: Lipolytic enzyme, G-D-S-L - Medicago truncatula (Barrel medic), partial (77%) TC268582 GMGI.071508 768733 771727 weakly similar to UniRef100_A7PMA0 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (89%) BE059369 Glycine _(—) max_release_2 770328 771326 Lipolytic enzyme = G-D-S-L [Medicago truncatula (Barrel medic)] BE329784 GMGI.071508 770783 771236 similar to UniRef100_Q1RU40 Cluster: Lipolytic enzyme = G-D-S-L; n = 1; Medicago truncatula|Rep: Lipolytic enzyme = G-D-S-L - Medicago truncatula (Barrel medic) = partial (27%) BE329784 Glycine _(—) max_release_2 770783 771288 Lipolytic enzyme = G-D-S- L [Medicago truncatula (Barrel medic)] TA68573_3847 Glycine _(—) max_release_2 773983 774836 Putative kinesin light chain [Oryza sativa (japonica cultivar-group)] TC259227 GMGI.071508 773983 774836 similar to UniRef100_A7PD12 Cluster: Chromosome chr17 scaffold_12 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr17 scaffold_12 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (13%) AI759741 Glycine _(—) max_release_2 774118 774822 Putative kinesin light chain [Oryza sativa (japonica cultivar-group)] asmbl_11876 Vigna _(—) unguiculata 774030 774978 NA TC139308 MTGI.071708 774935 775598 similar to UniRef100_A7PMA1 Cluster: Chromosome chr14 scaffold_21, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr14 scaffold_21, whole genome shotgun sequence - Vitis vinifera (Grape), partial (34%) AW186182 Glycine _(—) max_release_2 775276 775796 Similarity to kinesin light chain [Arabidopsis thaliana (Mouse-ear cress)] AW186182 GMGI.071508 775464 775796 similar to UniRef100_A7PD12 Cluster: Chromosome chr17 scaffold_12 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr17 scaffold_12 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (16%) BF010272 GMGI.071508 783671 784035 UniRef100_Q00K67 Cluster: Major surface antigen; n = 1; Hepatitis B virus|Rep: Major surface antigen - Hepatitis B virus (HBV) = partial (5%) TA54422_3847 Glycine _(—) max_release_2 783644 784982 Alcohol dehydrogenase superfamily = zinc- containing [Medicago truncatula (Barrel medic)] BI971258 Glycine _(—) max_release_2 783921 784926 Auxin-induced protein [Vigna radiata] CV542673 Phaseolus _(—) vulgaris _(—) {grave over ( )}{grave over ( )} 785346 Quinone oxidoreductase- release_2 like protein [Helianthus annuus (Common sunflower)] TC239445 GMGI.071508 783904 786356 similar to UniRef100_O23939 Cluster: Ripening-induced protein; n = 1; Fragaria vesca|Rep: Ripening- induced protein - Fragaria vesca (Woodland strawberry) = partial (84%) TA3037_3848 Glycine _(—) soja _(—) 784204 786191 Quinone oxidoreductase- release_2 like protein [Helianthus annuus (Common sunflower)] BG045149 Glycine _(—) soja _(—) 784943 785469 Quinone oxidoreductase release_2 [Fragaria ananassa (Strawberry)] TA54423_3847 Glycine _(—) max_release_2 784420 786354 Quinone oxidoreductase- like protein [Helianthus annuus (Common sunflower)] BG046280 Glycine _(—) soja _(—) 786163 786344 NA release_2 CA901808 Phaseolus _(—) coccineus _(—) 800890 801759 Alcohol dehydrogenase release_2 superfamily, zinc- containing [Medicago truncatula (Barrel medic)] TA14086_34305 Lotus _(—) japonicus _(—) 800932 801745 Alcohol dehydrogenase release_1 superfamily, zinc- containing [Medicago truncatula (Barrel medic)] TC23841 LJGI.070108 800932 801745 similar to UniRef100_Q43677 Cluster: Auxin-induced protein, n = 1, Vigna radiata|Rep: Auxin-induced protein - Vigna radiata, partial (40%) M0093116 SEQ. Listing 805373 805788 SEQ ID NO: 13 TC252650 GMGI.071508 805357 806601 similar to UniRef100_Q43677 Cluster: Auxin-induced protein; n = 1; Vigna radiata|Rep: Auxin-induced protein - Vigna radiata = partial (54%) BARC-039375-07304 ePCR&blat 805660 806532 Map3.0 SNP L/Gm19 cM: 3.4 TA65006_3847 Glycine _(—) max_release_2 805357 807089 Quinone oxidoreductase- like protein [Helianthus annuus (Common sunflower)] TA65005_3847 Glycine _(—) max_release_2 806611 807310 Alcohol dehydrogenase superfamily = zinc- containing [Medicago truncatula (Barrel medic)] TC274718 GMGI.071508 806611 807310 similar to UniRef100_Q43677 Cluster: Auxin-induced protein; n = 1; Vigna radiata|Rep: Auxin-induced protein - Vigna radiata = partial (30%) AW397551 Glycine _(—) max_release_2 811245 811796 Auxin-induced protein [Vigna radiata] Pvcon4580 Phaseolus _(—) vulgaris 811330 813524 UniRef100_Q43677 Auxin- induced protein n = 1 Tax = Vigna radiata RepID = Q43677_9FABA 1.00E−133 asmbl_11877 Vigna _(—) unguiculata 812523 812779 NA BE608172 Glycine _(—) max_release_2 821487 822389 Protein farnesyltransferase subunit beta [Pisum sativum (Garden pea)] BQ273477 Glycine _(—) max_release_2 821559 822383 NA TC246895 GMGI.071508 821516 822443 similar to UniRef100_Q04903 Cluster: Protein farnesyltransferase subunit beta; n = 1; Pisum sativum|Rep: Protein farnesyltransferase subunit beta - Pisum sativum (Garden pea) = partial (15%) TC241767 GMGI.071508 824186 828116 similar to UniRef100_Q7XHJ0 Cluster: Formate dehydrogenase; n = 1; Quercus robur|Rep: Formate dehydrogenase - Quercus robur (English oak) = partial (97%) TA40711_3847 Glycine _(—) max_release_2 824209 828372 Formate dehydrogenase [Quercus robur (English oak)] AI522957 Glycine _(—) max_release_2 826883 827087 Formate dehydrogenase [Quercus robur (English oak)] BG044450 Glycine _(—) soja _(—) 826544 827461 Formate dehydrogenase = release_2 mitochondrial precursor [Solanum tuberosum (Potato)] asmbl_11878 Vigna _(—) unguiculata 826586 827463 NA CA800817 Glycine _(—) soja _(—) 826705 827869 Formate dehydrogenase release_2 [Quercus robur (English oak)] TC240429 GMGI.071508 826957 828379 similar to UniRef100_Q9ZRI8 Cluster: Formate dehydrogenase = mitochondrial precursor; n = 1; Hordeum vulgare|Rep: Formate dehydrogenase = mitochondrial precursor - Hordeum vulgare (Barley) = partial (40%) AW350528 Glycine _(—) max_release_2 826986 828379 Formate dehydrogenase 1 = mitochondrial precursor [Oryza sativa (Rice)] BG882062 Glycine _(—) max_release_2 827372 828284 Formate dehydrogenase 1 = mitochondrial precursor [Oryza sativa (Rice)] BE347639 Glycine _(—) max_release_2 827443 828262 Formate dehydrogenase 1 = mitochondrial precursor [Oryza sativa (Rice)] CA782711 Glycine _(—) soja _(—) 827371 828357 Formate dehydrogenase 1 = release_2 mitochondrial precursor [Oryza sativa (Rice)] TA40821_3847 Glycine _(—) max_release_2 829640 832253 Formate dehydrogenase [Quercus robur (English oak)] BE330555 Glycine _(—) max_release_2 829875 832057 Formate dehydrogenase = mitochondrial precursor [Solanum tuberosum (Potato)] BU090495 Glycine _(—) max_release_2 829863 832082 Formate dehydrogenase [Quercus robur (English oak)] BG044406 Glycine _(—) soja _(—) 829915 832082 Formate dehydrogenase release_2 [Quercus robur (English oak)] AW508186 GMGI.071508 830914 831336 similar to UniRef100_A7PMA5 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (15%) M0129925 SEQ LISTING 830552 831704 SEQ ID NO: 14 AW508186 Glycine _(—) max_release_2 830914 831970 Formate dehydrogenase = mitochondrial precursor [Solatium tuberosum (Potato)] AW508145 Glycine _(—) max_release_2 830909 832061 Formate dehydrogenase [Quercus robur (English oak)] TA40373_3847 Glycine _(—) max_release_2 830863 832118 Formate dehydrogenase [Quercus robur (English oak)] AW397259 Glycine _(—) max_release_2 831219 832141 Formate dehydrogenase [Quercus robur (English oak)] TC261330 GMGI.071508 829795 833576 similar to UniRef100_Q7XHJ0 Cluster: Formate dehydrogenase; n = 1; Quercus robur|Rep: Formate dehydrogenase - Quercus robur (English oak) = partial (96%) TC249502 GMGI.071508 830866 832529 similar to UniRef100_Q7XHJ0 Cluster: Formate dehydrogenase; n = 1; Quercus robur|Rep: Formate dehydrogenase - Quercus robur (English oak) = partial (72%) TA40376_3847 Glycine _(—) max_release_2 830879 833356 Formate dehydrogenase [Quercus robur (English oak)] asmbl_11879 Vigna _(—) unguiculata 831735 833050 NA AW569072 GMGI.071508 832471 832890 similar to UniRef100_Q7XHJ0 Cluster: Formate dehydrogenase; n = 1; Quercus robur|Rep: Formate dehydrogenase - Quercus robur (English oak) = partial (9%) AW569072 Glycine _(—) max_release_2 832471 832929 Formate dehydrogenase [Quercus robur (English oak)] TA40339_3847 Glycine _(—) max_release_2 832130 833531 Formate dehydrogenase 1 = mitochondrial precursor [Oryza sativa (Rice)] TA5191_3885 Phaseolus _(—) vulgaris _(—) 832192 833517 Formate dehydrogenase release_2 [Quercus robur (English oak)] FD790937 Phaseolus _(—) vulgaris 833039 833412 UniRef100_A6N0B2 Mitochondrial formate dehydrogenase 1 (Fragment) n = 1 Tax = Oryza sativa Indica Group RepID = A6N0B2_ORYSI 3.00E−30 CA913454 Phaseolus _(—) coccineus _(—) 841331 841722 NA release_2 TA70199_3847 Glycine _(—) max_release_2 841305 841824 NA asmbl_11880 Vigna _(—) unguiculata 841326 841889 NA TA3611_3848 Glycine _(—) soja _(—) 841347 842640 Hypothetical protein release_2 OJ1593_C11.11 [Oryza sativa (japonica cultivar- group)] TA5381_34305 Lotus _(—) japonicus _(—) 841455 842700 Calcium homeostasis release_1 regulator CHoR1 [Solanum tuberosum (Potato)] TC20706 LJGI.070108 841455 842700 weakly similar to UniRef100_Q5QTN8 Cluster: Calcium homeostasis regulator CHoR1, n = 1, Solanum tuberosum|Rep: Calcium homeostasis regulator CHoR1 - Solanum tuberosum (Potato), partial (52%) Pvcon2378 Phaseolus _(—) vulgaris 841347 843522 UniRef100_A7PMA9 Chromosome chr14 scaffold_21, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7PMA9_VITVI 4.00E−94 TC252755 GMGI.071508 841305 843655 similar to UniRef100_A7PMA9 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (74%) EX305183 Phaseolus _(—) vulgaris 841682 843613 UniRef100_A7PMA9 Chromosome chr14 scaffold_21, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7PMA9_VITVI 1.00E−67 BI498351 GMGI.071508 844582 845168 NA TA66563_3847 Glycine _(—) max_release_2 844582 847078 Hypothetical protein [Ipomoea trifida (Morning glory)] TC247953 GMGI.071508 844582 847220 similar to UniRef100_A7Q5T8 Cluster: Chromosome chr14 scaffold_54 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_54 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (58%) TA3593_3848 Glycine _(—) soja _(—) 844668 847194 Hypothetical protein release_2 [Ipomoea trifida (Morning glory)] TA56324_3847 Glycine _(—) max_release_2 854425 856413 Similarity to intracellular protein [Arabidopsis thaliana (Mouse-ear cress)] TC235843 GMGI.071508 854425 856413 similar to UniRef100_A7PMB1 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (40%) CD406351 Glycine _(—) max_release_2 855627 856402 Similarity to intracellular protein [Arabidopsis thaliana (Mouse-ear cress)] TC276442 GMGI.071508 855627 856402 similar to UniRef100_A7PMB1 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (14%) TC273993 GMGI.071508 863632 864262 homologue to UniRef100_A7PMB2 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (26%) BU082700 Glycine _(—) max_release_2 863841 864449 Hypothetical protein OJ1126_B10.9 [Oryza sativa (japonica cultivar- group)] AW459960 Glycine _(—) max_release_2 863632 865288 Hypothetical protein F4P13.4 [Arabidopsis thaliana (Mouse-ear cress)] AL385435 MTGI.071708 863952 865397 homologue to UniRef100_A7PD25 Cluster: Chromosome chr17 scaffold_12, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr17 scaffold_12, whole genome shotgun sequence - Vitis vinifera (Grape), partial (37%) AI856244 GMGI.071508 864500 864958 UniRef100_A7PMB2 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (6%) asmbl_11881 Vigna _(—) unguiculata 863829 865710 NA TC238318 GMGI.071508 863970 865869 homologue to UniRef100_A7PMB2 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (34%) TA63907_3847 Glycine _(—) max_release_2 864500 865869 Hypothetical protein F4P13.4 [Arabidopsis thaliana (Mouse-ear cress)] BW598574 LJGI.070108 865265 865656 similar to UniRef100_Q8LES3 Cluster: Protein kinase, n = 1, Arabidopsis thaliana|Rep: Protein kinase - Arabidopsis thaliana (Mouse-ear cress), partial (9%) BW598574 Lotus _(—) japonicus _(—) 865265 865674 Protein kinase [Arabidopsis release_1 thaliana (Mouse-ear cress)] CD400016 Glycine _(—) max_release_2 870972 871184 NA CD399245 Glycine _(—) max_release_2 870876 871427 Putative Peptidyl-prolyl cis- trans isomerase = chloroplast [Oryza sativa (japonica cultivar-group)] TC242592 GMGI.071508 870943 872827 similar to UniRef100_A6MZC4 Cluster: Peptidyl-prolyl cis- trans isomerase; n = 2; Oryza sativa|Rep: Peptidyl-prolyl cis-trans isomerase - Oryza sativa subsp. indica (Rice) = partial (60%) CB543642 Phaseolus _(—) vulgaris _(—) 871229 872777 Peptidyl-prolyl cis-trans release_2 isomerase = chloroplast precursor [Spinacia oleracea (Spinach)] TA52959_3847 Glycine _(—) max_release_2 870943 873450 Poly(A) polymerase [Pisum sativum (Garden pea)] CB539263 Phaseolus _(—) vulgaris _(—) 871195 873325 Poly(A) polymerase [Pisum release_2 sativum (Garden pea)] Pvcon1578 Phaseolus _(—) vulgaris 870946 876143 UniRef100_O22636 Poly(A) polymerase n = 1 Tax = Pisum sativum RepID = O22636_PEA E−0 TA10487_34305 Lotus _(—) japonicus _(—) 873266 875963 Poly(A) polymerase [Pisum release_1 sativum (Garden pea)] TA6667_47247 Lotus _(—) corniculatus _(—) 873266 875963 Poly(A) polymerase related release_1 cluster TC34747 LJGI.070108 873266 875963 similar to UniRef100_O22636 Cluster: Poly(A) polymerase, n = 1, Pisum sativum|Rep: Poly(A) polymerase - Pisum sativum (Garden pea), partial (57%) BG363373 Glycine _(—) max_release_2 874357 874944 Poly(A) polymerase [Pisum sativum (Garden pea)] TC251420 GMGI.071508 874369 876078 similar to UniRef100_O22636 Cluster: Poly(A) polymerase; n = 1; Pisum sativum|Rep: Poly(A) polymerase - Pisum sativum (Garden pea) = partial (37%) CA901088 Phaseolus _(—) coccineus _(—) 874490 876191 Poly(A) polymerase [Pisum release_2 sativum (Garden pea)] asmbl_11882 Vigna _(—) unguiculata 886629 890018 NA TA68870_3847 Glycine _(—) max_release_2 886534 893419 Senescence-associated protein-like [Oryza sativa (japonica cultivar-group)] TC270337 GMGI.071508 886672 893419 weakly similar to UniRef100_A7PD28 Cluster: Chromosome chr17 scaffold_12 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr17 scaffold_12 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (86%) M0205537 SEQ. Listing 890458 890051 SEQ ID NO: 15 BM732054 Glycine _(—) max_release_2 899859 901015 NA BM732054 GMGI.071508 900006 901015 similar to UniRef100_Q04TD2 Cluster: MviN-related protein; n = 1; Leptospira borgpetersenii serovar Hardjo-bovis JB197|Rep: = partial (2%) toGm13 DAGchainer 816170 1014875 Ks0.1202 M0202715 SEQ. Listing 921233 921630 SEQ ID NO: 16 TA46168_3847 Glycine _(—) max_release_2 921047 924660 Homeodomain leucine zipper protein HDZ3 [Phaseolus vulgaris (Kidney bean) (French bean)] TC260016 GMGI.071508 921056 924739 homologue to UniRef100_Q93XA3 Cluster: Homeodomain leucine zipper protein HDZ3; n = 1; Phaseolus vulgaris|Rep: Homeodomain leucine zipper protein HDZ3 - Phaseolus vulgaris (Kidney bean) (French bean) = complete Pvcon1101 Phaseolus _(—) vulgaris 921086 924758 UniRef100_Q93XA3 Homeodomain leucine zipper protein HDZ3 (Fragment) n = 1 Tax = Phaseolus vulgaris RepID = Q93XA3_PHAVU 1.00E−124 TA3604_3885 Phaseolus _(—) vulgaris _(—) 921111 924754 Homeodomain leucine release_2 zipper protein HDZ3 [Phaseolus vulgaris (Kidney bean) (French bean)] asmbl_11883 Vigna _(—) unguiculata 921538 924758 NA BG041631 Glycine _(—) soja _(—) 923015 923340 Homeobox-leucine zipper release_2 protein HAT5 [Arabidopsis thaliana (Mouse-ear cress)] AV421688 LJGI.070108 923118 924180 similar to UniRef100_Q93XA3 Cluster: Homeodomain leucine zipper protein HDZ3, n = 1, Phaseolus vulgaris|Rep: Homeodomain leucine zipper protein HDZ3 - Phaseolus vulgaris (Kidney bean) (French bean), partial (25%) TC235979 GMGI.071508 923000 924768 similar to UniRef100_Q93XA3 Cluster: Homeodomain leucine zipper protein HDZ3; n = 1; Phaseolus vulgaris|Rep: Homeodomain leucine zipper protein HDZ3 - Phaseolus vulgaris (Kidney bean) (French bean) = partial (86%) TA46165_3847 Glycine _(—) max_release_2 923000 924779 Homeodomain leucine zipper protein HDZ3 [Phaseolus vulgaris (Kidney bean) (French bean)] AW351287 Glycine _(—) max_release_2 923128 924720 Homeodomain leucine zipper protein HDZ3 [Phaseolus vulgaris (Kidney bean) (French bean)] CA785782 Glycine _(—) soja _(—) 925713 925880 NA release_2 Pvcon8364 Phaseolus _(—) vulgaris 925735 926609 UniRef100_A7PMB7 Chromosome chr14 scaffold_21, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7PMB7_VITVI 1.00E−27 BE248998 MTGI.071708 926978 927524 similar to UniRef100_Q7F8S7 Cluster: PHD finger-like protein, n = 1, Oryza sativa Japonica Group|Rep: PHD finger-like protein - Oryza sativa subsp. japonica (Rice), partial (4%) TC35470 LJGI.070108 928423 929804 similar to UniRef100_A7PMB8 Cluster: Chromosome chr14 scaffold_21, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr14 scaffold_21, whole genome shotgun sequence - Vitis vinifera (Grape), partial (9%) TA11035_34305 Lotus _(—) japonicus _(—) 928423 929825 PHD finger-like protein release_1 [Oryza sativa (japonica cultivar-group)] CA911004 Phaseolus _(—) coccineus _(—) 934882 939256 T13O15.10 protein release_2 [Arabidopsis thaliana (Mouse-ear cress)] AI856399 GMGI.071508 937577 938041 NA AI856399 Glycine _(—) max_release_2 937577 938106 NA AW348703 Glycine _(—) max_release_2 963043 963750 NA TC276191 GMGI.071508 963049 964044 weakly similar to UniRef100_A7PZY3 Cluster: Chromosome chr8 scaffold_41 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr8 scaffold_41 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (21%) BQ628183 Glycine _(—) max_release_2 963625 964044 NA BQ080193 Glycine _(—) max_release_2 963695 967475 NA TA52645_3847 Glycine _(—) max_release_2 963720 967461 NA TC256882 GMGI.071508 963774 967475 weakly similar to UniRef100_A7PZY3 Cluster: Chromosome chr8 scaffold_41 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr8 scaffold_41 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (45%) BG156825 Glycine _(—) max_release_2 971121 971284 NA BG156825 GMGI.071508 971125 971284 NA BU545761 Glycine _(—) max_release_2 971300 971901 NA BU550718 Glycine _(—) max_release_2 971255 973578 NA TA72701_3847 Glycine _(—) max_release_2 972120 972806 NA TC271942 GMGI.071508 972201 972806 NA TC269989 GMGI.071508 971255 973827 similar to UniRef100_A7P2M9 Cluster: Chromosome chr1 scaffold_5 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr1 scaffold_5 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (63%) BI317782 Glycine _(—) max_release_2 971510 973827 NA BI893512 Glycine _(—) max_release_2 971537 973848 NA BI893512 GMGI.071508 971671 973848 similar to UniRef100_A7P2M9 Cluster: Chromosome chr1 scaffold_5 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr1 scaffold_5 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (54%) CO985587 Glycine _(—) max_release_2 974859 976255 Putative GTP-binding membrane protein LepA [Oryza sativa (japonica cultivar-group)] AW596868 Glycine _(—) max_release_2 976346 976856 NA AW596868 GMGI.071508 976412 976856 similar to UniRef100_A2Q5T1 Cluster: Tetratricopeptide- like helical; n = 1; Medicago truncatula|Rep: Tetratricopeptide-like helical - Medicago truncatula (Barrel medic) = partial (5%) CA901672 Phaseolus _(—) coccineus _(—) 983905 984264 Aldehyde dehydrogenase 1 release_2 precursor [Lotus corniculatus (Bird's-foot trefoil) WmFPC_Contig4169 899736 1068750 NA FE898889 Phaseolus _(—) vulgaris 983908 984989 UniRef100_A7PD33 Chromosome chr17 scaffold_12, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7PD33_VITVI 2.00E−79 TC273361 GMGI.071508 984396 986122 similar to UniRef100_P93344 Cluster: Aldehyde dehydrogenase; n = 1; Nicotiana tabacum|Rep: Aldehyde dehydrogenase - Nicotiana tabacum (Common tobacco) = partial (37%) BE473475 Glycine _(—) max_release_2 984960 986122 Aldehyde dehydrogenase [Nicotiana tabacum (Common tobacco)] CV539672 Phaseolus _(—) vulgaris 985959 987101 UniRef100_P93344 Aldehyde dehydrogenase (NAD+) n = 1 Tax = Nicotiana tabacum RepID = P93344_TOBAC 7.00E−50 AV410805 LJGI.070108 987592 987888 similar to UniRef100_A7PMC7 Cluster: Chromosome chr14 scaffold_21, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr14 scaffold_21, whole genome shotgun sequence - Vitis vinifera (Grape), partial (6%) TC265505 GMGI.071508 1011306 1012664 similar to UniRef100_A7PMD1 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (28%) TA51641_3847 Glycine _(—) max_release_2 1011306 1013783 Putative high-affinity potassium transporter protein 1 [Nicotiana tabacum (Common tobacco)] CB540416 Phaseolus _(—) vulgaris 1012333 1013531 UniRef100_A7PMD1 Chromosome chr14 scaffold_21, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7PMD1_VITVI 5.00E−97 BM891067 GMGI.071508 1012675 1013617 similar to UniRef100_A7PMD1 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (22%) TC131883 MTGI.071708 1012665 1014070 similar to UniRef100_A7PMD1 Cluster: Chromosome chr14 scaffold_21, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr14 scaffold_21, whole genome shotgun sequence - Vitis vinifera (Grape), partial (34%) asmbl_11884 Vigna _(—) unguiculata 1012674 1014123 NA BE330787 Glycine _(—) max_release_2 1013888 1014305 Putative high-affinity potassium transporter protein [Phytolacca esculenta (Food pokeberry)] FD792954 Phaseolus _(—) vulgaris 1013779 1014573 UniRef100_A7PMD1 Chromosome chr14 scaffold_21, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7PMD1_VITVI 3.00E−57 TC244134 GMGI.071508 1014004 1014793 similar to UniRef100_A7PMD1 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (15%) TA51642_3847 Glycine _(—) max_release_2 1013926 1014875 Putative high-affinity potassium transporter 1 [Nicotiana rustica (Aztec tobacco)] TC242106 GMGI.071508 1013926 1014875 similar to UniRef100_A7PMD1 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (15%) BI970123 Glycine _(—) max_release_2 1014128 1014721 Putative potassium transporter HAK1p [Mesembryanthemum crystallinum (Common ice plant)] BQ080303 Glycine _(—) max_release_2 1018604 1019142 NA TC270109 GMGI.071508 1018604 1019142 weakly similar to UniRef100_UPI0000196D3 9 Cluster: NHL repeat- containing protein; n = 1; Arabidopsis thaliana|Rep: NHL repeat-containing protein - Arabidopsis thaliana = partial (4%) BQ080219 Glycine _(—) max_release_2 1018604 1019579 NA TA62145_3847 Glycine _(—) max_release_2 1021347 1023221 NA TC245123 GMGI.071508 1021347 1023221 similar to UniRef100_A7PMD2 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (31%) asmbl_11885 Vigna _(—) unguiculata 1022417 1022510 NA CA784724 Glycine _(—) max_release_2 1046117 1047384 NA CA784724 GMGI.071508 1046400 1047384 similar to UniRef100_A7Q2E7 Cluster: Chromosome chr1 scaffold_46 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr1 scaffold_46 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (17%) Pvcon4015 Phaseolus _(—) vulgaris 1047011 1048610 UniRef100_A5ATC1 Putative uncharacterized protein n = 1 Tax = Vitis vinifera RepID = A5ATC1_VITVI 1.00E−146 BQ742289 Glycine _(—) max_release_2 1048650 1048767 NA BF068315 GMGI.071508 1057203 1057316 similar to UniRef100_Q8MIG1 Cluster: Skinkine; n = 1; Sus scrofa|Rep: Skinkine - Sus scrofa (Pig) = partial (12%) BF068315 Glycine _(—) max_release_2 1057203 1057506 NA BU083500 GMGI.071508 1058026 1058431 UniRef100_Q2R023 Cluster: Expressed protein; n = 1; Oryza sativa Japonica Group|Rep: Expressed protein - Oryza sativa = partial (2%) TA74227_3847 Glycine _(—) max_release_2 1058026 1059408 NA BI423963 GMGI.071508 1058432 1059275 similar to UniRef100_Q2QDD6 Cluster: Nodulin-like protein; n = 1; Gossypium hirsutum|Rep: Nodulin-like protein - Gossypium hirsutum (Upland cotton) (Gossypium mexicanum) = partial (22%) TC237120 GMGI.071508 1063015 1063972 UniRef100_Q39819 Cluster: Hsp22.3; n = 1; Glycine max|Rep: Hsp22.3 - Glycine max (Soybean) = complete CA802234 Glycine _(—) soja _(—) 1061477 1067499 Similarity to nodulin release_2 [Arabidopsis thaliana (Mouse-ear cress)] BI425574 GMGI.071508 1065519 1066854 weakly similar to UniRef100_A7PMD8 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (21%) BI425574 Glycine _(—) max_release_2 1065519 1066940 Hypothetical protein [Medicago truncatula (Barrel medic)] AU251786 LJGI.070108 1066790 1067424 weakly similar to UniRef100_A7Q2G7 Cluster: Chromosome chr1 scaffold_46, whole genome shotgun sequence, n = 1, Vitis vinifera|Rep: Chromosome chr1 scaffold_46, whole genome shotgun sequence - Vitis vinifera (Grape), partial (7%) Pvcon8451 Phaseolus _(—) vulgaris 1065511 1068752 UniRef100_A7PMD8 Chromosome chr14 scaffold_21, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7PMD8_VITVI 7.00E−91 TC260900 GMGI.071508 1065796 1069134 weakly similar to UniRef100_A7PMD8 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (41%) TA63020_3847 Glycine _(—) max_release_2 1067436 1069134 NA CA783703 Glycine _(—) soja _(—) 1068257 1068879 NA release_2 TA58065_3847 Glycine _(—) max_release_2 1074998 1076541 AT3g28050/MMG15_6 [Arabidopsis thaliana (Mouse-ear cress)] TC251785 GMGI.071508 1074998 1076541 similar to UniRef100_Q8L9I2 Cluster: Nodulin MtN21- like protein; n = 1; Arabidopsis thaliana|Rep: Nodulin MtN21-like protein - Arabidopsis thaliana (Mouse-ear cress) = partial (16%) CB280623 Phaseolus _(—) vulgaris _(—) 1075036 1076540 AT3g28050/MMG15_6 release_2 [Arabidopsis thaliana (Mouse-ear cress)] EH043320 Arachis _(—) stenosperma _(—) 1075056 1077422 Cluster: Hypothetical release_5 protein, n = 1, Medicago truncatula|Rep: Hypothetical protein - Medicago truncatula (Barrel medic) asmbl_11886 Vigna _(—) unguiculata 1075036 1077585 NA BQ094260 Glycine _(—) max_release_2 1075548 1077551 Nodulin-like protein [Arabidopsis thaliana (Mouse-ear cress)] BF598290 Glycine _(—) soja _(—) 1075557 1077593 Nodulin-like protein release_2 [Arabidopsis thaliana (Mouse-ear cress)] Pvcon6314 Phaseolus _(—) vulgaris 1075036 1078733 UniRef100_A7PMD8 Chromosome chr14 scaffold_21, whole genome shotgun sequence n = 1 Tax = Vitis vinifera RepID = A7PMD8_VITVI 1.00E−105 TA58064_3847 Glycine _(—) max_release_2 1075337 1079189 AT3g28050/MMG15_6 [Arabidopsis thaliana (Mouse-ear cress)] TC255833 GMGI.071508 1075337 1079189 weakly similar to UniRef100_A7PMD8 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (64%) BG042956 Glycine _(—) soja _(—) 1078885 1079014 NA release_2 TC263589 GMGI.071508 1086875 1091139 similar to UniRef100_A7PME0 Cluster: Chromosome chr14 scaffold_21 = whole genome shotgun sequence; n = 1; Vitis vinifera|Rep: Chromosome chr14 scaffold_21 = whole genome shotgun sequence - Vitis vinifera (Grape) = partial (35%) TA50577_3847 Glycine _(—) max_release_2 1086875 1094082 Alpha-dioxygenase [Pisum sativum (Garden pea)] asmbl_11887 Vigna _(—) unguiculata 1089135 1092345 NA CA410123 Lupinus _(—) albus _(—) 1092182 1092694 Alpha-dioxygenase [Pisum release_2 sativum (Garden pea) Pvcon4974 Phaseolus _(—) vulgaris 1091225 1093836 UniRef100_Q5GQ66 Alpha-dioxygenase n = 1 Tax = Pisum sativum RepID = Q5GQ66_PEA E−0 TC243973 GMGI.071508 1091177 1094141 similar to UniRef100_Q5GQ66 Cluster: Alpha- dioxygenase; n = 1; Pisum sativum|Rep: Alpha- dioxygenase - Pisum sativum (Garden pea) = partial (61%) asmbl_11888 Vigna _(—) unguiculata 1092518 1093829 NA M0206286 SEQ. Listing 1209562 1210392 SEQ ID NO: 17 M0206054 SEQ. Listing 1465522 1465187 SEQ ID NO: 18 M0205375 SEQ. Listing 2010060 2009541 SEQ ID NO: 19 toGm13 DAGchainer 1046081 4647877 Ks0.2059 NA Glyma1 1 50600000 NA

Sequences for the genes provided above can be obtained from the World Wide Web (or Internet) using the identifiers provided in Column 1 (Locus/Display Name) or Column 5 (ADDITIONAL LOCUS INFORMATION) from the following internet locations: “soybase.org” (described in Grant et al., Nucleic Acids Research, 2010, Vol. 38, Database issue D843-D846) or soybase.org/gbrowse/cgi-bin/gbrowse/gmax1.01/(see Hyten D L, Choi I-Y, Song Q, Specht J E, Carter T E et al. (2010) A high density integrated genetic linkage map of soybean and the development of a 1,536 Universal Soy Linkage Panel for QTL mapping. Crop Science 50:960-968. (Crop Science); and Hyten D L, Cannon S B, Song Q, Weeks N, Fickus E W et al. (2010) High-throughput SNP discovery through deep resequencing of a reduced representation library to anchor and orient scaffolds in the soybean whole genome sequence. BMC Genomics 11(1): 38);

“phytozome.net” or “phytozome.net/cgi-bin/gbrowse/soybean/?name=Gm09”; “www.plantgdb.org” or “plantgdb.org/GmGDB/ (Assembly version Glyrnal.170 (April 2009)”; and, “ncbi.nlm.nih.gov/sites/entrez” and subsites “ncbi.nlm.nih.gov/nucest”, “ncbi.nlm.nih.gov/dbEST”, “ncbi.nlm.nih.gov/genbank/”, “.ncbi.nlm.nih.gov/sites/genome”, “ncbi.nlm.nih.gov/unigene”, and “ncbi.nlm.nih.gov/UniGene/UGOrg.cgi?TAXID=3847”. 

We claim:
 1. A method of identifying a soybean plant that comprises a genotype associated with dicamba tolerance and reproductive tolerance to glyphosate, comprising: detecting in a soybean plant an allele in at least one genetic locus associated with dicamba tolerance and reproductive tolerance to glyphosate, wherein the genetic locus is in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12), and denoting that said plant comprises a genotype associated with dicamba tolerance.
 2. The method of claim 1, wherein said method further comprises the step of selecting said denoted plant from a population of plants.
 3. The method of claim 1, wherein said plant comprises a transgene that confers resistance to dicamba and/or a transgene that confers resistance to glyphosate.
 4. The method of claim 3, wherein said soybean plant or progeny thereof is exposed to a dosage of dicamba sufficient to cause a deleterious effect in a susceptible variety comprising the transgene and/or is exposed to a dosage of glyphosate sufficient to cause sterility in a susceptible variety comprising the transgene(s).
 5. The method of claim 2, wherein a plant that exhibits dicamba tolerance and/or reproductive tolerance to glyphosate is selected.
 6. The method of claim 1, wherein said genotype associated with a dicamba tolerance comprises at least one polymorphic allele of at least one marker in a first sub-region of said linkage group L region that is flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6) and/or at least one polymorphic allele of at least one marker in a second sub-region of said linkage group L region that is flanked by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12) and/or at least one polymorphic allele of at least one marker in a third sub-region of said linkage group L region that is flanked by loci BU55345 (SEQ ID NO:7) and M0114388 (SEQ ID NO:8).
 7. The method of any one of claims 1-6, wherein said genotype associated with dicamba tolerance comprises at least one polymorphic allele of at least one marker in said linkage group L region selected from the group consisting of a TT allele M0205350 (SEQ ID NO: 10), a TT allele of M0101742 (SEQ ID NO: 5), a CC allele of M0102027 (SEQ ID NO: 11), and an AA allele of NGMAX008197032 (SEQ ID NO:52).
 8. A method for obtaining a soybean plant comprising in its genome at least one dicamba tolerance locus, compromising the steps of: (a) genotyping a plurality of soybean plants with respect to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12); and (b) selecting a soybean plant comprising in its genome at least one genetic locus comprising a genotype associated with dicamba tolerance.
 9. The method of claim 8, wherein said genotype associated with dicamba tolerance comprises at least one polymorphic allele of at least one marker in a first sub-region of said linkage group L region flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6) and/or at least one polymorphic allele of at least one marker in a second sub-region of said linkage group L region that is flanked by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12) and/or at least one polymorphic allele of at least one marker in a third sub-region of said linkage group L region that is flanked by loci BU55345 (SEQ ID NO:7) and M0114388 (SEQ ID NO:8).
 10. The method of claim 8, wherein said genotype associated with dicamba tolerance comprises at least one polymorphic allele of at least one marker in said first linkage group L region, said first sub-region, or said second sub-region, wherein said marker is selected from the group consisting of a TT allele M0205350 (SEQ ID NO: 10), a TT allele of M0101742 (SEQ ID NO: 5), a CC allele of M0102027 (SEQ ID NO: 11), and an AA allele of NGMAX008197032 (SEQ ID NO:52).
 11. The method of claim 8, wherein said plurality of soybean plants comprises a population that is obtained by: i) crossing a parent plant comprising at least one dicamba tolerance locus with a parent plant comprising at least one dicamba sensitivity locus; or, ii) obtaining seed or progeny from a parental plant segregating for at least one dicamba tolerance locus.
 12. The method of claim 8, wherein said population contains plants that comprise a transgene that confers resistance to dicamba and/or a transgene that confers resistance to glyphosate.
 13. The method of any one of claims 8-12, further comprising the step of assaying for the presence of at least one additional marker, wherein said additional marker is either linked or unlinked to said linkage group L genomic region.
 14. The method of any one of claims 8-12, wherein said plurality of soybean plants, said soybean plant, and/or progeny thereof are exposed to a dosage of dicamba sufficient to cause a deleterious effect in a susceptible variety comprising the transgene and/or is exposed to a dosage of glyphosate sufficient to cause sterility in a susceptible variety comprising the transgene.
 15. The method of any one of claims 8-12, wherein a plant that exhibits dicamba tolerance and/or reproductive tolerance to glyphosate is selected.
 16. A method for producing a soybean plant comprising in its genome at least one introgressed dicamba tolerance locus comprising the steps of: (a) crossing a first soybean plant with a dicamba tolerance locus with a second soybean plant comprising: a dicamba sensitivity locus in a first linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6) and/or at least one polymorphic allele of at least one marker in a second sub-region of said linkage group L region that is flanked by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12) and/or at least one polymorphic allele of at least one marker in a third sub-region of said linkage group L region that is flanked by loci BU55345 (SEQ ID NO:7) and M0114388 (SEQ ID NO:8) and at least one linked polymorphic locus not present in said first soybean plant to obtain a population segregating for the dicamba tolerance loci and said linked polymorphic locus; (b) detecting at least two polymorphic nucleic acids in at least one soybean plant from said population, wherein at least one of said polymorphic nucleic acids is located in said first linkage group L region and/or said second linkage group L region and wherein at least one of said polymorphic amino acids is a linked polymorphic locus not present in said first soybean plant; and (c) selecting a soybean plant comprising a genotype associated with dicamba tolerance and at least one linked marker found in said second soybean plant comprising a dicamba sensitivity locus but not in said first soybean plant, thereby obtaining a soybean plant comprising in its genome at least one introgressed dicamba tolerance locus.
 17. The method of claim 16, wherein at least one of said first or said second soybean plants comprises a transgene that confers resistance to dicamba and/or a transgene that confers resistance to glyphosate.
 18. The method of claim 17, wherein said population, said selected soybean plant, and/or progeny of selected soybean plant is exposed to a dosage of dicamba sufficient to cause a deleterious effect in a susceptible variety comprising the transgene and/or is exposed to a dosage of glyphosate sufficient to cause sterility in a susceptible variety comprising the transgene.
 19. The method of claim 16, wherein said polymorphic nucleic acid detected in step (b) is detected with at least one marker selected from the group consisting of M0205350 (SEQ ID NO: 10), M0101742 (SEQ ID NO: 5), M0102027 (SEQ ID NO: 11), and NGMAX008197032 (SEQ ID NO:52).
 20. The method of claim 16, wherein said polymorphic nucleic acid detected in step (b) comprises a TT allele of M0205350 (SEQ ID NO: 10), a TT allele of M0101742 (SEQ ID NO: 5), a CC allele of M0102027 (SEQ ID NO: 11), and an AA allele of NGMAX008197032 (SEQ ID NO:52).
 21. The method of claim 20, wherein said polymorphic nucleic acid detected in step (b) is detected with marker M0205350 (SEQ ID NO: 10), M0102027 (SEQ ID NO: 11), or marker NGMAX008197032 (SEQ ID NO:52).
 22. The method of claim 20, wherein said polymorphic nucleic acids are detected with marker M0101742 (SEQ ID NO: 5).
 23. The method of any one of claims 16-22, wherein said linked polymorphic locus is detected with a genotypic marker, a phenotypic marker, or both.
 24. The method of claim 23, wherein said linked polymorphic locus is detected with a marker that is located within about 1000, 500, 100, 40, 20, 10, or 5 kilobases (Kb) of said dicamba tolerance locus.
 25. The method of claim 24, wherein said linked polymorphic locus is detected with at least one marker selected from the group consisting of asmbl_11856 (SEQ ID NO: 1), TC122822 (SEQ ID NO: 2), BI967232 (SEQ ID NO: 3), M0205537 (SEQ ID NO: 15), M0202715 (SEQ ID NO: 16), M0206286 (SEQ ID NO: 17), M0206054 (SEQ ID NO:18), and M0205375 (SEQ ID NO: 19).
 26. A transgenic soybean plant comprising introgressed linkage group L regions comprising at least one polymorphic allele of at least one marker in a first sub-region of said linkage group L region that flanked by loci M0205928 (SEQ ID NO: 4) and M0129138 (SEQ ID NO: 6) and/or at least one polymorphic allele of at least one marker in a second sub-region of said linkage group L region that is flanked by loci BU551363 (SEQ ID NO: 9) and BU765955 (SEQ ID NO: 12) and/or at least one polymorphic allele of at least one marker in a third sub-region of said linkage group L region that is flanked by loci BU55345 (SEQ ID NO:7) and M0114388 (SEQ ID NO:8), wherein said polymorphic alleles are associated with dicamba tolerance and/or reproductive tolerance to glyphosate, and wherein said plant comprises a transgene that confers resistance to dicamba.
 27. The transgenic plant of claim 26, wherein said polymorphic alleles comprise a TT allele of M0205350 (SEQ ID NO: 10), a TT allele of M0101742 (SEQ ID NO: 5), a CC allele of M0102027 (SEQ ID NO: 11), and an AA allele of NGMAX008197032 (SEQ ID NO:52).
 28. The transgenic plant of claim 26, wherein said plant exhibits dicamba tolerance.
 29. The transgenic plant of claim 26, wherein said plant further comprises a transgene that confers resistance to glyphosate and exhibits reproductive tolerance to glyphosate.
 30. The transgenic plant of any one of claims 26-29, wherein said plant further comprises at least one transgene conferring resistance to a herbicide selected from the group consisting of a 2,4-D, glufosinate, bromoxynil, synthetic auxins other than 2,4-D, acetolactate synthase (ALS), acetyl CoA carboxylase (ACCase), hydroxyphenyl pyruvate dioxygenase (HPPD), and a sulfonylurea herbicide and/or at least one transgene selected from the group of transgenes conferring insect resistance, nematode resistance, fungal resistance, an improvement in seed oil quantity, an improvement in seed oil quality, abiotic stress resistance, and intrinsic yield increases.
 31. A method of identifying a transgenic soybean plant that comprises a genotype associated with reproductive tolerance to glyphosate, the method comprising: (a) scoring at least one transgenic plant in a population of transgenic soybean plants that had been exposed to dicamba for dicamba tolerance, said plants having a transgene that confers resistance to dicamba; and, (b) selecting a transgenic plant that exhibits dicamba tolerance, thereby identifying a transgenic soybean plant that comprises a genotype associated with reproductive tolerance to glyphosate.
 32. The method of claim 31, wherein said population is segregating for to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12) that is associated with dicamba tolerance.
 33. The method of claim 31, wherein said method further comprises genotyping the selected soybean plant with respect to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12).
 34. The method of any one of claims 31-33, wherein said selected transgenic plant further comprises a transgene that confers resistance to glyphosate and wherein said selected transgenic plant or progeny thereof is scored for reproductive tolerance to glyphosate following exposure to glyphosate.
 36. The method of any one of claims 31-33, further comprising exposing said population of transgenic soybean plants to dicamba.
 37. The method of any one of claims 31-33, wherein dicamba tolerance is scored by determining a reduction in malformation when compared to a dicamba sensitive transgenic plant that comprises said transgene that confers resistance to dicamba.
 38. A method of identifying a transgenic soybean plant that comprises a genotype associated with tolerance to dicamba, comprising: (a) scoring at least one plant in a population of transgenic soybean plants that had been exposed to glyphosate for reproductive tolerance to glyphosate, wherein said plants comprise a transgene that confers resistance to glyphosate; and, (b) selecting a transgenic plant that exhibits reproductive tolerance to glyphosate, thereby identifying a transgenic soybean plant that comprises a genotype associated with dicamba tolerance.
 39. The method of claim 38, wherein said population is segregating for to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12) that is associated with dicamba tolerance.
 40. The method of claim 38, wherein said method further comprises genotyping the selected soybean plant with respect to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12).
 41. The method of any one of claims 38-40, wherein said selected transgenic plant further comprises a transgene that confers resistance to dicamba and wherein said selected transgenic plant or progeny thereof is scored for tolerance to dicamba following exposure to dicamba.
 42. The method of any one of claims 38-40, further comprising exposing said population of transgenic soybean plants to glyphosate.
 43. The method of any one of claims 38-40, wherein glyphosate reproductive tolerance is scored by determining a reduction in sterility when compared to a transgenic plant that exhibits glyphosate reproductive sensitivity and comprises said transgene that confers resistance to glyphosate.
 44. A method of obtaining a transgenic soybean plant that comprises a genotype associated with reproductive tolerance to glyphosate, the method comprising: (a) exposing a population of transgenic soybean plants to dicamba, wherein said plants have a transgene that confers resistance to dicamba; (b) observing dicamba tolerance exhibited by one or more soybean plants following exposure to dicamba; and, (c) selecting a transgenic plant that exhibits dicamba tolerance, thereby obtaining a transgenic soybean plant that comprises a genotype associated with reproductive tolerance to glyphosate.
 45. The method of claim 44, wherein said population is segregating for to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12) that is associated with dicamba tolerance.
 46. The method of claim 44, wherein said method further comprises genotyping the selected soybean plant with respect to at least one genetic locus in a linkage group L genomic region flanked by loci M0205928 (SEQ ID NO: 4) and BU765955 (SEQ ID NO: 12).
 47. The method of any one of claims 44-46, wherein said selected transgenic plant further comprises a transgene that confers resistance to glyphosate and wherein said selected transgenic plant or progeny thereof is scored for reproductive tolerance to glyphosate following exposure to glyphosate.
 48. The method of any one of claims 44-46, wherein dicamba tolerance is scored by determining a reduction in malformation when compared to a dicamba sensitive transgenic plant that comprises said transgene that confers resistance to dicamba. 